LOCUS       sequence-12039-e        4216 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 4216)
  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 4216)
  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-TK-Acceptor
FEATURES             Location/Qualifiers
     source          1..4216
                     /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)."
     CDS             521..1324
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(584..603)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1194..1213
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1331..1379
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    complement(1380..1413)
                     /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(1440..1456)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1441..1457)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     repeat_region   1479..1619
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1479..1608
                     /label=AAV2 ITR
     rep_origin      1694..2149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1781..1800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1991..2012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2166..2185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2285..2307
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2345..2363)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2431..2535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2536..2604,2605..3396)
                     /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(2754..2773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3567..4155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4056..4075
                     /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 cgaagttatc
      241 tcgagcagtg tggttttgca agaggaagca aaaagcctct ccacccaggc ctggaatgtt
      301 tccacccaat gtcgagcagt gtggttttgc aagaggaagc aaaaagcctc tccacccagg
      361 cctggaatgt ttccacccaa tgtcgagcaa accccgccca gcgtcttgtc attggcgaat
      421 tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg
      481 acgcgtgtgg cctcgaacac cgagcgaccc tgcagccaat atgggatcgg ccattgaaca
      541 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
      601 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
      661 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
      721 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
      781 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
      841 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
      901 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
      961 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     1021 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     1081 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     1141 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     1201 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     1261 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     1321 ctgaggggat cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     1381 taacttcgta tagcatacat tatacgaagt tatccgcggc ctagggatat caagcttatc
     1441 gataccgtcg acctcgaggg gccccggccg gcggccgcag gaacccctag tgatggagtt
     1501 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     1561 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     1621 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     1681 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1741 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1801 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1861 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     1921 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1981 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     2041 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2101 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     2161 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2221 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2281 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2341 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2401 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2521 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2581 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2641 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2701 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2761 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2821 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2881 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2941 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3001 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3061 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3121 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3181 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3241 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3301 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3361 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3421 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3481 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3541 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3601 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3661 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3721 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3781 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3841 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3901 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3961 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4021 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4081 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4141 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4201 ccttttgctc acatgt
//
