LOCUS       sequence-430554-        5009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5009)
  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 5009)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2008 Nov . 36(19):e127."
COMMENT     Sequence Label: pAAV-SEPT-Acceptor
FEATURES             Location/Qualifiers
     source          1..5009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    complement(206..239)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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    complement(2158..2191)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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..2412
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2474..3062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2554..2573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3233..4024,4025..4093))
                     /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     3856..3875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4094..4198)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4266..4284
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4322..4344)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4444..4463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4480..4935)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4638)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4829..4848
                     /label=F1ori-R
                     /note="F1 origin, reverse 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 gggcggcctc agtgagcgag cgagcgcgca
     2401 gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2881 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3181 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3301 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3361 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3421 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3481 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3541 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3601 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3661 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3721 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3781 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3841 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3901 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3961 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4021 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4081 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4201 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4261 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4321 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4381 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4441 atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt
     4501 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa
     4561 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag
     4621 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4681 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     4741 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4801 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg
     4861 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4921 cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg
     4981 cgtaaggaga aaataccgca tcaggcgcc
//
