LOCUS Exported 5020 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAV-SEPT-Acceptor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5020) AUTHORS Kim JS, Bonifant C, Bunz F, Lane WS, Waldman T TITLE Epitope tagging of endogenous genes in diverse human cell lines. JOURNAL Nucleic Acids Res. 2008 Nov . 36(19):e127. PUBMED 18784188 REFERENCE 2 (bases 1 to 5020) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5020 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR primer_bind 188..204 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 188..204 /label=pBluescriptSK /note="For pBluescript vector" protein_bind 206..239 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." intron 277..506 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 489..508 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 635..1097 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(691..708) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 918..937 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 1111..1905 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1165..1184) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1775..1794 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1927..2061 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1951..1970) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2005..2024 /label=SV40pA-R /note="SV40 polyA, reverse primer" protein_bind 2158..2191 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(2244..2260) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2245..2261) /label=pBluescriptKS /note="For pBluescript vector" repeat_region 2283..2423 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2283..2412 /label=AAV2 ITR rep_origin 2498..2953 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2585..2604) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2795..2816 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2970..2989) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3089..3111 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3149..3167) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3235..3339 /gene="bla" /label=AmpR promoter CDS 3340..4200 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3558..3577) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4371..4959 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4860..4879 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca ccggtggcgc gcctgatcaa cgcgtgctag 181 cgagctctct agaactagtg gatccataac ttcgtatagc atacattata cgaagttatg 241 gaattaattc gctgtctgcg agggccagct gttggggtga gtactccctc tcaaaagcgg 301 gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg atattcacct 361 ggcccgcggt gatgcctttg agggtggccg cgtccatctg gtcagaaaag acaatctttt 421 tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc atacacttga gtgacaatga 481 catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc aggtcgagcg 541 tcgagaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt 601 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct 661 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga 721 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga 781 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac 841 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag 901 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc 961 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg 1021 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt 1081 gaaaaacacg atgataagct tgccacaacc atggctgaac aagatggatt gcacgcaggt 1141 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 1201 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 1261 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg 1321 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 1381 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 1441 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 1501 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 1561 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 1621 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 1681 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 1741 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 1801 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 1861 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 1921 tcgggggatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg 1981 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt 2041 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag 2101 ggggaggtgt gggaggtttt ttctaggggg ccccggcgag gccgggatga gttgggaata 2161 acttcgtata gcatacatta tacgaagtta tccaaatttt aaggtaccac tgtgcatatg 2221 tatgaattcg atatcaagct tatcgatacc gtcgacctcg aggggccccg gccggcggcc 2281 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 2341 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga 2401 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg 2461 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta 2521 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 2581 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 2641 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 2701 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt 2761 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 2821 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc 2881 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta 2941 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 3001 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 3061 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 3121 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 3181 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 3241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 3301 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 3361 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 3421 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 3481 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 3541 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 3601 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 3661 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 3721 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 3781 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 3841 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 3901 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 3961 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 4021 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 4081 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 4141 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 4201 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 4261 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 4321 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 4381 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 4441 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 4501 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 4561 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 4621 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 4681 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 4741 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 4801 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 4861 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 4921 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 4981 ttacggttcc tggccttttg ctggcctttt gctcacatgt //