LOCUS P4HB-pGEc2-DEST 6733 bp DNA circular 26-JAN-2013 FEATURES Location/Qualifiers misc_feature complement(6211..6340) /note="3'-ITR" misc_feature complement(5909..6123) /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 5361..5902 /note="WPRE" misc_recomb complement(2974..3001) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 4540..4562 /note="attB2" misc_feature 5340..5345 /note="BstB1 site" misc_feature 4563..5258 /note="Super GFP" misc_feature 5259..5309 /note="AviTag" misc_feature 5310..5333 /note="His Tag" PCR_frame 3002..4540 /note="Reading Frame" /set="Set 1" source 3006..4517 /dnas_title="P4HB-pGEc2-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:88571 IMAGE:5443660" /tissue_type="Skin, melanoma, amelanotic" /clone_lib="NIH_MGC_41" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 3006..4517 /dnas_title="P4HB" /gene="P4HB" /gene_synonym="DSI" /gene_synonym="GIT" /gene_synonym="P4Hbeta" /gene_synonym="PDI" /gene_synonym="PDIA1" /gene_synonym="PHDB" /gene_synonym="PO4HB" /gene_synonym="PROHB" /db_xref="GeneID:5034" /db_xref="HGNC:8548" /db_xref="MIM:176790" CDS 3006..4517 /dnas_title="P4HB" /gene="P4HB" /gene_synonym="DSI" /gene_synonym="GIT" /gene_synonym="P4Hbeta" /gene_synonym="PDI" /gene_synonym="PDIA1" /gene_synonym="PHDB" /gene_synonym="PO4HB" /gene_synonym="PROHB" /codon_start=1 /product="prolyl 4-hydroxylase, beta polypeptide" /protein_id="AAH71892.1" /db_xref="GI:48735337" /db_xref="GeneID:5034" /db_xref="HGNC:8548" /db_xref="MIM:176790" /translation="MLRRALLCLAVAALVRADAPEEEDHVLVLRKSNFAEALAAHKYL LVEFYAPWCGHCKALAPEYAKAAGKLKAEGSEIRLAKVDATEESDLAQQYGVRGYPTI KFFRNGDTASPKEYTAGREADDIVNWLKKRTGPAATTLPDGAAAESLVESSEVAVIGF FKDVESDSAKQFLQAAEAIDDIPFGITSNSDVFSKYQLDKDGVVLFKKFDEGRNNFEG EVTKENLLDFIKHNQLPLVIEFTEQTAPKIFGGEIKTHILLFLPKSVSDYDGKLSNFK TAAESFKGKILFIFIDSDHTDNQRILEFFGLKKEECPAVRLITLEEEMTKYKPESEEL TAERITEFCHRFLEGKIKPHLMSQELPEDWDKQPVKVLVGKNFEDVAFDEKKNVFVEF YAPWCGHCKQLAPIWDKLGETYKDHENIVIAKMDSTANEVEAVKVHSFPTLKFFPASA DRTVIDYNGERTLDGFKKFLESGGQDGAGDDDDLEDLEEAEEPDMEEDDDQKAVKDEL " misc_feature 3006..4517 /note="P4HB coding region" misc_feature 4518..4538 /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..4547) /dnas_title="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /primer="AAAGCTGGGTTTAGCAGTGAATGGTCCG" /set="Set 1" /pair="Pair 1" /current=1 /original="ACCACTTTGTACAAGAAAGCTGGGTTTAGCAGTGAATGGTCCG" PCR_product complement(2992..4547) /pair="Pair 1" /set="Set 1" /current=1 /score="1.000" vector join(4548..6733,1..2991) /source="pGEc2-DEST" /type="Destination Vector" clone 2992..4547 /source="P4HB-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 TCGCCatgct gcgccgcgct ctgctgtgcc tggccgtggc cgccctggtg cgcgccgacg 3061 cccccgagga ggaggaccac gtcctggtgc tgcggaaaag caacttcgcg gaggcgctgg 3121 cggcccacaa gtacctgctg gtggagttct atgccccttg gtgtggccac tgcaaggctc 3181 tggcccctga gtatgccaaa gccgctggga agctgaaggc agaaggttcc gagatcaggt 3241 tggccaaggt ggacgccacg gaggagtctg acctggccca gcagtacggc gtgcgcggct 3301 atcccaccat caagttcttc aggaatggag acacggcttc ccccaaggaa tatacagctg 3361 gcagagaggc tgatgacatc gtgaactggc tgaagaagcg cacgggcccg gctgccacca 3421 ccctgcctga cggcgcagct gcagagtcct tggtggagtc cagcgaggtg gctgtcatcg 3481 gcttcttcaa ggacgtggag tcggactctg ccaagcagtt tttgcaggca gcagaggcca 3541 tcgatgacat accatttggg atcacttcca acagtgacgt gttctccaaa taccagctcg 3601 acaaagatgg ggttgtcctc tttaagaagt ttgatgaagg ccggaacaac tttgaagggg 3661 aggtcaccaa ggagaacctg ctggacttta tcaaacacaa ccagctgccc cttgtcatcg 3721 agttcaccga gcagacagcc ccgaagattt ttggaggtga aatcaagact cacatcctgc 3781 tgttcttgcc caagagtgtg tctgactatg acggcaaact gagcaacttc aaaacagcag 3841 ccgagagctt caagggcaag atcctgttca tcttcatcga cagcgaccac accgacaacc 3901 agcgcatcct cgagttcttt ggcctgaaga aggaagagtg cccggccgtg cgcctcatca 3961 ccctggagga ggagatgacc aagtacaagc ccgaatcgga ggagctgacg gcagagagga 4021 tcacagagtt ctgccaccgc ttcctggagg gcaaaatcaa gccccacctg atgagccagg 4081 agctgccgga ggactgggac aagcagcctg tcaaggtgct tgttgggaag aactttgaag 4141 acgtggcttt tgatgagaaa aaaaacgtct ttgtggagtt ctatgcccca tggtgtggtc 4201 actgcaaaca gttggctccc atttgggata aactgggaga gacgtacaag gaccatgaga 4261 acatcgtcat cgccaagatg gactcgactg ccaacgaggt ggaggccgtc aaagtgcaca 4321 gcttccccac actcaagttc tttcctgcca gtgccgacag gacggtcatt gattacaacg 4381 gggaacgcac gctggatggt tttaagaaat tcctggagag cggtggccag gatggggcag 4441 gggatgatga cgatctcgag gacctggaag aagcagagga gccagacatg gaggaagacg 4501 atgatcagaa agctgtggaa aacttgtact ttcaaggctC CAGCTTTCTT GTACAAAGTG 4561 GTATGAGCAA GGGCGAGGAA CTGTTCACCG GCGTGGTGCC CATCCTGGTG GAACTGGATG 4621 GCGACGTGAA CGGCCACAAG TTCTCTGTGC GGGGAGAGGG CGAAGGCGAC GCCACAAATG 4681 GCAAGCTGAC CCTGAAGTTC ATCTGCACCA CCGGCAAGCT GCCCGTGCCT TGGCCTACCC 4741 TCGTGACCAC ACTGACCTAC GGCGTGCAGT GCTTCAGCAG ATACCCCGAC CATATGAAGC 4801 GGCACGACTT CTTCAAGAGC GCCATGCCCG AGGGCTACGT GCAGGAACGG ACCATCAGCT 4861 TCAAGGACGA CGGCACCTAC AAGACCAGAG CCGAAGTGAA GTTCGAGGGC GACACCCTCG 4921 TGAACCGGAT CGAGCTGAAG GGCATCGATT TCAAAGAGGA CGGCAACATC CTGGGCCACA 4981 AGCTGGAGTA CAACTTCAAC AGCCACAACG TGTACATCAC CGCCGACAAG CAGAAGAACG 5041 GCATCAAGGC CAACTTCAAG ATCCGGCACA ACGTGGAAGA TGGCAGCGTG CAGCTGGCCG 5101 ACCACTACCA GCAGAACACC CCCATCGGAG ATGGCCCCGT GCTGCTGCCC GACAACCACT 5161 ACCTGAGCAC CCAGAGCGTG CTGAGCAAGG ACCCCAACGA GAAGCGGGAC CACATGGTGC 5221 TGCTGGAATT CGTGACCGCC GCTGGCATCA CCCACGGCAT GAGCGGCCTG AACGACATCT 5281 TCGAGGCCCA GAAAATCGAG TGGCACGAGC ACCACCATCA TCACCATCAC CACTGATGAT 5341 TCGAAAAATA ATAAGGATCC AATCAACCTC TGGATTACAA AATTTGTGAA AGATTGACTG 5401 GTATTCTTAA CTATGTTGCT CCTTTTACGC TATGTGGATA CGCTGCTTTA ATGCCTTTGT 5461 ATCATGCTAT TGCTTCCCGT ATGGCTTTCA TTTTCTCCTC CTTGTATAAA TCCTGGTTGC 5521 TGTCTCTTTA TGAGGAGTTG TGGCCCGTTG TCAGGCAACG TGGCGTGGTG TGCACTGTGT 5581 TTGCTGACGC AACCCCCACT GGTTGGGGCA TTGCCACCAC CTGTCAGCTC CTTTCCGGGA 5641 CTTTCGCTTT CCCCCTCCCT ATTGCCACGG CGGAACTCAT CGCCGCCTGC CTTGCCCGCT 5701 GCTGGACAGG GGCTCGGCTG TTGGGCACTG ACAATTCCGT GGTGTTGTCG GGGAAGCTGA 5761 CGTCCTTTCC ATGGCTGCTC GCCTGTGTTG CCACCTGGAT TCTGCGCGGG ACGTCCTTCT 5821 GCTACGTCCC TTCGGCCCTC AATCCAGCGG ACCTTCCTTC CCGCGGCCTG CTGCCGGCTC 5881 TGCGGCCTCT TCCGCGTCTT CGAGATCTGC CTCGACTGTG CCTTCTAGTT GCCAGCCATC 5941 TGTTGTTTGC CCCTCCCCCG TGCCTTCCTT GACCCTGGAA GGTGCCACTC CCACTGTCCT 6001 TTCCTAATAA AATGAGGAAA TTGCATCGCA TTGTCTGAGT AGGTGTCATT CTATTCTGGG 6061 GGGTGGGGTG GGGCAGGACA GCAAGGGGGA GGATTGGGAA GACAATAGCA GGCATGCTGG 6121 GGACTCGAGT TAAGGGCGAA TTGCCGATTA GGATCTTCCT AGAGCATGGC TACGTAGATA 6181 AGTAGCATGG CGGGTTAATC ATTAACTACA AGGAACCCCT AGTGATGGAG TTGGCCACTC 6241 CCTCTCTGCG CGCTCGCTCG CTCACTGAGG CCGGGCGACC AAAGGTCGCC CGACGCCCGG 6301 GCTTTGCCCG GGCGGCCTCA GTGAGCGAGC GAGCGCGCAG CCTTAATTAA GAGCTCATGG 6361 CGCGCCTAGG CCTTGACGGC CTTCCGCCAA TTCGCCCTAT AGTGAGTCGT ATTACGTCGC 6421 GCTCACTGGC CGTCGTTTTA CAACGTCGTG ACTGGGAAAA CCCTGGCGTT ACCCAACTTA 6481 ATCGCCTTGC AGCACATCCC CCTTTCGCCA GCTGGCGTAA TAGCGAAGAG GCCCGCACCG 6541 AAACGCCCTT CCCAACAGTT GCGCAGCCTG AATGGCGAAT GGGAGCGCCC TGTAGCGGCC 6601 ACTCAACCCT ATCTCGGTCT ATTCTTTTGA TTTATAAGGG ATTTTGCCGA TTTCGGCCTA 6661 TTGGTTAAAA AATGAGCTGA TTTAACAAAA ATTTAACGCG AATTTTAACA AAATATTAAC 6721 GCTTACAATT TAG //