LOCUS 40924_1444 4566 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector JSC, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4566) AUTHORS Sepulveda J, Shoemaker CB. TITLE Design and testing of PCR primers for the construction of scFv libraries representing the immunoglobulin repertoire of rats JOURNAL J. Immunol. Methods 332 (1-2), 92-102 (2008) PUBMED 18242637 REFERENCE 2 (bases 1 to 4566) AUTHORS Shoemaker CB, Sepulveda J. TITLE Direct Submission JOURNAL Submitted (22-AUG-2007) Biomedical Science, Tufts Cummings School of Veterinary Medicine, 200 Westboro Rd., North Grafton, MA 01536, USA REFERENCE 3 (bases 1 to 4566) TITLE Direct Submission REFERENCE 4 (bases 1 to 4566) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Immunol. Methods 332 (1-2), 92-102 (2008)" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (22-AUG-2007) Biomedical Science, Tufts Cummings School of Veterinary Medicine, 200 Westboro Rd., North Grafton, MA 01536, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..4566 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 330..351 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 366..396 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 404..420 /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." sig_peptide 443..508 /label=pelB signal sequence /note="leader peptide for secretion" misc_feature 508..593 /label=multi-cloning site /note="multi-cloning site" misc_feature 593..631 /label=E-tag /note="E-tag" CDS 641..1852 /codon_start=1 /label=M13 gene III /note="pIII" /translation="TVESCLAKPHTENSFTNVWKDDKTLDRYANYEGCLWNATGVVVCT GDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGGTKPPEYGDTPIPGYTYINPL DGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQGALTVYTGTVTQGTDPVKTY YQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQGQSSDLPQPPVNAGGGSGGG SGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKMANANKGAMTENADENALQSD AKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGDNSPLMNNFR QYLPSLPQSVECRPYVFGAGKPYEFSIDCDKINLFRGVFAFLLYVATFMYVFSTFANIL RNKES" primer_bind complement(1862..1878) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 2091..2546 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2828..2932 /label=AmpR promoter CDS 2933..3790 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3964..4552 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt 61 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 121 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg 181 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agggtacccg ataaaagcgg 241 cttcctgaca ggaggccgtt ttgttttgca gcccacctct ggcacgacag gtttcccgac 301 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc 361 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa 421 ttgaattcag gaggaattta aaatgaaata cctattgcct acggcagccg ctggattgtt 481 attactcgcg gcccaaccgg ccatggccgg cccgggagcg gccgctggtg gaggcggttc 541 aggcggaggt ggctctggcg gtggcggatc ggcgcgccag gcctgcacta gtggtgcgcc 601 ggtgccgtat ccggatccgc tggaaccgcg tgccgcatag actgttgaaa gttgtttagc 661 aaaacctcat acagaaaatt catttactaa cgtctggaaa gacgacaaaa ctttagatcg 721 ttacgctaac tatgagggct gtctgtggaa tgctacaggc gttgtggttt gtactggtga 781 cgaaactcag tgttacggta catgggttcc tattgggctt gctatccctg aaaatgaggg 841 tggtggctct gagggtggcg gttctgaggg tggcggttct gagggtggcg gtactaaacc 901 tcctgagtac ggtgatacac ctattccggg ctatacttat atcaaccctc tcgacggcac 961 ttatccgcct ggtactgagc aaaaccccgc taatcctaat ccttctcttg aggagtctca 1021 gcctcttaat actttcatgt ttcagaataa taggttccga aataggcagg gtgcattaac 1081 tgtttatacg ggcactgtta ctcaaggcac tgaccccgtt aaaacttatt accagtacac 1141 tcctgtatca tcaaaagcca tgtatgacgc ttactggaac ggtaaattca gagactgcgc 1201 tttccattct ggctttaatg aggatccatt cgtttgtgaa tatcaaggcc aatcgtctga 1261 cctgcctcaa cctcctgtca atgctggcgg cggctctggt ggtggttctg gtggcggctc 1321 tgagggtggc ggctctgagg gtggcggttc tgagggtggc ggctctgagg gtggcggttc 1381 cggtggcggc tccggttccg gtgattttga ttatgaaaaa atggcaaacg ctaataaggg 1441 ggctatgacc gaaaatgccg atgaaaacgc gctacagtct gacgctaaag gcaaacttga 1501 ttctgtcgct actgattacg gtgctgctat cgatggtttc attggtgacg tttccggcct 1561 tgctaatggt aatggtgcta ctggtgattt tgctggctct aattcccaaa tggctcaagt 1621 cggtgacggt gataattcac ctttaatgaa taatttccgt caatatttac cttctttgcc 1681 tcagtcggtt gaatgtcgcc cttatgtctt tggcgctggt aaaccatatg aattttctat 1741 tgattgtgac aaaataaact tattccgtgg tgtctttgcg tttcttttat atgttgccac 1801 ctttatgtat gtattttcga cgtttgctaa catactgcgt aataaggagt cttaagaatt 1861 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 1921 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 1981 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc 2041 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg 2101 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 2161 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 2221 ttttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 2281 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg 2341 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 2401 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt 2461 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 2521 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc 2581 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 2641 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 2701 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt 2761 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 2821 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 2881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 2941 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 3001 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 3061 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 3121 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 3181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 3241 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 3301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 3361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 3421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 3481 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 3541 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 3601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 3661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 3721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 3781 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 3841 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 3901 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 3961 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 4021 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 4081 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 4141 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 4201 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 4261 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 4321 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 4381 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 4441 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 4501 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 4561 caacgc //