LOCUS SPAM1-pGEc2-DEST 6688 bp DNA circular 25-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6166..6295) /note="3'-ITR" misc_feature complement(5864..6078) /note="BGH pA" rep_origin 1137..1804 /dnas_title="Col E1 origin" /label=Col E1 origin /vntifkey="33" CDS 129..989 /dnas_title="AmpR" /label=AmpR /vntifkey="4" misc_feature 1998..2127 /note="5'ITR" misc_feature 2208..2790 /note="CMV Promoter" misc_feature 2828..2960 /note="Artificial Intron" misc_feature 5316..5857 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 4495..4517 /note="attB2" misc_feature 5295..5300 /note="BstB1 site" misc_feature 4518..5213 /note="Super GFP" misc_feature 5214..5264 /note="AviTag" misc_feature 5265..5288 /note="His Tag" PCR_frame 3002..4495 /note="Reading Frame" /set="Set 1" source 3006..4472 /dnas_title="SPAM1-pGEc2-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:26532 IMAGE:4838230" /tissue_type="Testis" /clone_lib="NIH_MGC_97" /lab_host="DH10B" /note="Vector: pBluescriptR" gene 3006..4472 /dnas_title="SPAM1" /gene="SPAM1" /gene_synonym="HYA1" /gene_synonym="HYAL1" /gene_synonym="HYAL3" /gene_synonym="MGC26532" /gene_synonym="PH-20" /gene_synonym="PH20" /gene_synonym="SPAG15" /db_xref="GeneID:6677" /db_xref="HGNC:11217" /db_xref="MIM:600930" CDS 3006..4472 /dnas_title="sperm adhesion molecule 1 (PH-20 hyaluronidase, zona pellucida binding)" /gene="SPAM1" /gene_synonym="HYA1" /gene_synonym="HYAL1" /gene_synonym="HYAL3" /gene_synonym="MGC26532" /gene_synonym="PH-20" /gene_synonym="PH20" /gene_synonym="SPAG15" /codon_start=1 /product="sperm adhesion molecule 1 (PH-20 hyaluronidase, zona pellucida binding)" /protein_id="AAH26163.1" /db_xref="GI:20072878" /db_xref="GeneID:6677" /db_xref="HGNC:11217" /db_xref="MIM:600930" /translation="MGVLKFKHIFFRSFVKSSGVSQIVFTFLLIPCCLTLNFRAPPVI PNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYI DSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKP KDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGY YLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATL YVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGAS GIVIWGTLSIMRSMKSCLLLDNYMETILNPYIINVTLAAKMCSQVLCQEQGVCIRKNW NSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVK DTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLSATMFIWRLEVWDQGISRI GFF" misc_feature 3006..4472 /note="SPAM1 coding region" misc_difference 3587..3587 /dnas_title="SPAM1" /gene="SPAM1" /gene_synonym="HYA1" /gene_synonym="HYAL1" /gene_synonym="HYAL3" /gene_synonym="MGC26532" /gene_synonym="PH-20" /gene_synonym="PH20" /gene_synonym="SPAG15" /note="'A' in cDNA is 'G' in the human genome; no amino acid change. The chimpanzee genome agrees with the human genomic sequence and not the cDNA." misc_feature 4473..4493 /note="TEV site" PCR_primer 2992..3001 /dnas_title="GTACAAAAAAGCAGGCTCATGGAACAAGGACAGCTCTTCC" /primer="CAGCTCTTCC" /set="Set 1" /pair="Pair 1" /current=1 /original="GTACAAAAAAGCAGGCTCATGGAACAAGGACAGCTCTTCC" PCR_primer complement(3002..4502) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..4502) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(4503..6688,1..2991) /source="pGEc2-DEST" /type="Destination Vector" clone 2992..4502 /source="SPAM1-C-term-TEVfusion-pDONR221" /type="Entry Clone" ORIGIN 1 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT 61 CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA 121 GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT 181 GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT 241 TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT 301 TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG 361 TATTATCCCG TATTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA 421 ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA 481 GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA 541 CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA 601 CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA 661 CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA 721 CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC 781 TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC 841 GTGGTTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG 901 TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA 961 TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT 1021 AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA 1081 ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG 1141 AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA 1201 CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT 1261 TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTTCTT CTAGTGTAGC 1321 CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA 1381 TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA 1441 GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC 1501 CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CTATGAGAAA 1561 GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA 1621 CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG 1681 GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC 1741 TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG 1801 CTCATTAGGC ACCCCAGGCT TTACCCGAAC GACCGAGCGC AGCGAGTCAG TGAGCGAGGA 1861 AGCGGAGAGC GCCCAATACG CAAGGAAACA GCTATGACCA TGTTAATGCA GCTGGCACGA 1921 CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGGAAGGCC CATGAGGCCA GTTAATTAAG 1981 AGGTACCTTA ATTAAGGCTG CGCGCTCGCT CGCTCACTGA GGCCGCCCGG GCAAAGCCCG 2041 GGCGTCGGGC GACCTTTGGT CGCCCGGCCT CAGTGAGCGA GCGAGCGCGC AGAGAGGGAG 2101 TGGCCAACTC CATCACTAGG GGTTCCTTGT AGTTAATGAT TAACCCGCCA TGCTACTTAT 2161 CTACGTAGCC ATGCTCTAGG AAGATCGCAA TTCGCCCTTA AGCTAGCTAG TTATTAATAG 2221 TAATCAATTA CGGGGTCATT AGTTCATAGC CCATATATGG AGTTCCGCGT TACATAACTT 2281 ACGGTAAATG GCCCGCCTGG CTGACCGCCC AACGACCCCC GCCCATTGAC GTCAATAATG 2341 ACGTATGTTC CCATAGTAAC GCCAATAGGG ACTTTCCATT GACGTCAATG GGTGGAGTAT 2401 TTACGGTAAA CTGCCCACTT GGCAGTACAT CAAGTGTATC ATATGCCAAG TACGCCCCCT 2461 ATTGACGTCA ATGACGGTAA ATGGCCCGCC TGGCATTATG CCCAGTACAT GACCTTATGG 2521 GACTTTCCTA CTTGGCAGTA CATCTACGTA TTAGTCATCG CTATTACCAT GGTGATGCGG 2581 TTTTGGCAGT ACATCAATGG GCGTGGATAG CGGTTTGACT CACGGGGATT TCCAAGTCTC 2641 CACCCCATTG ACGTCAATGG GAGTTTGTTT TGGCACCAAA ATCAACGGGA CTTTCCAAAA 2701 TGTCGTAACA ACTCCGCCCC ATTGACGCAA ATGGGCGGTA GGCGTGTACG GTGGGAGGTC 2761 TATATAAGCA GAGCTGGTTT AGTGAACCGT CAGATCCTGC AGAAGTTGGT CGTGAGGCAC 2821 TGGGCAGGTA AGTATCAAGG TTACAAGACA GGTTTAAGGA GACCAATAGA AACTGGGCTT 2881 GTCGAGACAG AGAAGACTCT TGCGTTTCTG ATAGGCACCT ATTGGTCTTA CTGACATCCA 2941 CTTTGCCTTT CTCTCCACAG GTGTCCAGCG GCCATCACAA GTTTGTACAA AAAAGCAGGC 3001 TCGCCatggg agtgctaaaa ttcaagcaca tctttttcag aagctttgtt aaatcaagtg 3061 gagtatccca gatagttttc accttccttc tgattccatg ttgcttgact ctgaatttca 3121 gagcacctcc tgttattcca aatgtgcctt tcctctgggc ctggaatgcc ccaagtgaat 3181 tttgtcttgg aaaatttgat gagccactag atatgagcct cttctctttc ataggaagcc 3241 cccgaataaa cgccaccggg caaggtgtta caatatttta tgttgataga cttggctact 3301 atccttacat agattcaatc acaggagtaa ctgtgaatgg aggaatcccc cagaagattt 3361 ccttacaaga ccatctggac aaagctaaga aagacattac attttatatg ccagtagaca 3421 atttgggaat ggctgttatt gactgggaag aatggagacc cacttgggca agaaactgga 3481 aacctaaaga tgtttacaag aataggtcta ttgaattggt tcagcaacaa aatgtacaac 3541 ttagtctcac agaggccact gagaaagcaa aacaagaatt tgaaaaagca gggaaggatt 3601 tcctggtaga gactataaaa ttgggaaaat tacttcggcc aaatcacttg tggggttatt 3661 atctttttcc ggattgttac aaccatcact ataagaaacc cggttacaat ggaagttgct 3721 tcaatgtaga aataaaaaga aatgatgatc tcagctggtt gtggaatgaa agcactgctc 3781 tttacccatc catttatttg aacactcagc agtctcctgt agctgctaca ctctatgtgc 3841 gcaatcgagt tcgggaagcc atcagagttt ccaaaatacc tgatgcaaaa agtccacttc 3901 cggtttttgc atatacccgc atagttttta ctgatcaagt tttgaaattc ctttctcaag 3961 atgaacttgt gtatacattt ggcgaaactg ttgctctggg tgcttctgga attgtaatat 4021 ggggaaccct cagtataatg cgaagtatga aatcttgctt gctcctagac aattacatgg 4081 agactatact gaatccttac ataatcaacg tcacactagc agccaaaatg tgtagccaag 4141 tgctttgcca ggagcaagga gtgtgtataa ggaaaaactg gaattcaagt gactatcttc 4201 acctcaaccc agataatttt gctattcaac ttgagaaagg tggaaagttc acagtacgtg 4261 gaaaaccgac acttgaagac ctggagcaat tttctgaaaa attttattgc agctgttata 4321 gcaccttgag ttgtaaggag aaagctgatg taaaagacac tgatgctgtt gatgtgtgta 4381 ttgctgatgg tgtctgtata gatgcttttc taaaacctcc catggagaca gaagaacctc 4441 aaattttcta caatgcttca ccctccacac tagaaaactt gtactttcaa ggctCCAGCT 4501 TTCTTGTACA AAGTGGTATG AGCAAGGGCG AGGAACTGTT CACCGGCGTG GTGCCCATCC 4561 TGGTGGAACT GGATGGCGAC GTGAACGGCC ACAAGTTCTC TGTGCGGGGA GAGGGCGAAG 4621 GCGACGCCAC AAATGGCAAG CTGACCCTGA AGTTCATCTG CACCACCGGC AAGCTGCCCG 4681 TGCCTTGGCC TACCCTCGTG ACCACACTGA CCTACGGCGT GCAGTGCTTC AGCAGATACC 4741 CCGACCATAT GAAGCGGCAC GACTTCTTCA AGAGCGCCAT GCCCGAGGGC TACGTGCAGG 4801 AACGGACCAT CAGCTTCAAG GACGACGGCA CCTACAAGAC CAGAGCCGAA GTGAAGTTCG 4861 AGGGCGACAC CCTCGTGAAC CGGATCGAGC TGAAGGGCAT CGATTTCAAA GAGGACGGCA 4921 ACATCCTGGG CCACAAGCTG GAGTACAACT TCAACAGCCA CAACGTGTAC ATCACCGCCG 4981 ACAAGCAGAA GAACGGCATC AAGGCCAACT TCAAGATCCG GCACAACGTG GAAGATGGCA 5041 GCGTGCAGCT GGCCGACCAC TACCAGCAGA ACACCCCCAT CGGAGATGGC CCCGTGCTGC 5101 TGCCCGACAA CCACTACCTG AGCACCCAGA GCGTGCTGAG CAAGGACCCC AACGAGAAGC 5161 GGGACCACAT GGTGCTGCTG GAATTCGTGA CCGCCGCTGG CATCACCCAC GGCATGAGCG 5221 GCCTGAACGA CATCTTCGAG GCCCAGAAAA TCGAGTGGCA CGAGCACCAC CATCATCACC 5281 ATCACCACTG ATGATTCGAA AAATAATAAG GATCCAATCA ACCTCTGGAT TACAAAATTT 5341 GTGAAAGATT GACTGGTATT CTTAACTATG TTGCTCCTTT TACGCTATGT GGATACGCTG 5401 CTTTAATGCC TTTGTATCAT GCTATTGCTT CCCGTATGGC TTTCATTTTC TCCTCCTTGT 5461 ATAAATCCTG GTTGCTGTCT CTTTATGAGG AGTTGTGGCC CGTTGTCAGG CAACGTGGCG 5521 TGGTGTGCAC TGTGTTTGCT GACGCAACCC CCACTGGTTG GGGCATTGCC ACCACCTGTC 5581 AGCTCCTTTC CGGGACTTTC GCTTTCCCCC TCCCTATTGC CACGGCGGAA CTCATCGCCG 5641 CCTGCCTTGC CCGCTGCTGG ACAGGGGCTC GGCTGTTGGG CACTGACAAT TCCGTGGTGT 5701 TGTCGGGGAA GCTGACGTCC TTTCCATGGC TGCTCGCCTG TGTTGCCACC TGGATTCTGC 5761 GCGGGACGTC CTTCTGCTAC GTCCCTTCGG CCCTCAATCC AGCGGACCTT CCTTCCCGCG 5821 GCCTGCTGCC GGCTCTGCGG CCTCTTCCGC GTCTTCGAGA TCTGCCTCGA CTGTGCCTTC 5881 TAGTTGCCAG CCATCTGTTG TTTGCCCCTC CCCCGTGCCT TCCTTGACCC TGGAAGGTGC 5941 CACTCCCACT GTCCTTTCCT AATAAAATGA GGAAATTGCA TCGCATTGTC TGAGTAGGTG 6001 TCATTCTATT CTGGGGGGTG GGGTGGGGCA GGACAGCAAG GGGGAGGATT GGGAAGACAA 6061 TAGCAGGCAT GCTGGGGACT CGAGTTAAGG GCGAATTGCC GATTAGGATC TTCCTAGAGC 6121 ATGGCTACGT AGATAAGTAG CATGGCGGGT TAATCATTAA CTACAAGGAA CCCCTAGTGA 6181 TGGAGTTGGC CACTCCCTCT CTGCGCGCTC GCTCGCTCAC TGAGGCCGGG CGACCAAAGG 6241 TCGCCCGACG CCCGGGCTTT GCCCGGGCGG CCTCAGTGAG CGAGCGAGCG CGCAGCCTTA 6301 ATTAAGAGCT CATGGCGCGC CTAGGCCTTG ACGGCCTTCC GCCAATTCGC CCTATAGTGA 6361 GTCGTATTAC GTCGCGCTCA CTGGCCGTCG TTTTACAACG TCGTGACTGG GAAAACCCTG 6421 GCGTTACCCA ACTTAATCGC CTTGCAGCAC ATCCCCCTTT CGCCAGCTGG CGTAATAGCG 6481 AAGAGGCCCG CACCGAAACG CCCTTCCCAA CAGTTGCGCA GCCTGAATGG CGAATGGGAG 6541 CGCCCTGTAG CGGCCACTCA ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGGGATTTT 6601 GCCGATTTCG GCCTATTGGT TAAAAAATGA GCTGATTTAA CAAAAATTTA ACGCGAATTT 6661 TAACAAAATA TTAACGCTTA CAATTTAG //