LOCUS       sequence-14382-e        3429 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 3429)
  AUTHORS   Dmitriev PV, Vassetzky YS
  TITLE     A set of vectors for introduction of antibiotic resistance genes by 
            in vitro Cre-mediated recombination.
  JOURNAL   BMC Res Notes. 2008 Dec 23;1:135. doi: 10.1186/1756-0500-1-135.
  PUBMED    19105832
REFERENCE   2  (bases 1 to 3429)
  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: "BMC Res 
            Notes."; date: "2008-12-23"; volume: "1:135. doi"; pages: " 
            10.1186/1756-0500-1-135"
COMMENT     Sequence Label: pINS-Neo
FEATURES             Location/Qualifiers
     source          1..3429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..362
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(415..448)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    506..554
                     /note="synthetic polyadenylation signal"
     CDS             complement(618..1412)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1463..1820)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1477..1612
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1629..1820
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1777..1797
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2015..2033)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2029..2076
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2280..3074)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 aattctgtca gccgttaagt gttcctgtgt cactgaaaat tgctttgaga ggctctaagg
       61 gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaagcttt
      121 aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt
      181 tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag
      241 cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca
      301 tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt
      361 actatcaaca ggttgaactg ctgatcaaca gatcctctac gcggccgcgg taccataact
      421 tcgtatagca tacattatac gaagttatct ggaattcccc gggctcgaga acatatggcc
      481 atggggatcc ttatcgctat cgattcacac aaaaaaccaa cacacagatg taatgaaaat
      541 aaagatattt tattgcggcc atcgtgatgg caggttgggc gtcgcttggt cggtcatttc
      601 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
      661 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
      721 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
      781 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
      841 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
      901 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
      961 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     1021 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     1081 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     1141 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     1201 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     1261 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     1321 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     1381 gaacctgcgt gcaatccatc ttgttcaatc atggtggctc tagccttaag ttcgagactg
     1441 ttgtgtcaga agaatcaagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     1501 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     1561 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     1621 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     1681 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     1741 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     1801 gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag
     1861 gccatggtgc tgcgcagatc cgcgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1921 aaaataccgc atcaggaaat tgtaagcgtt aattcgactc tagaagatcc ggctgctaac
     1981 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2041 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     2101 tatcccgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     2161 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     2221 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     2281 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     2341 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     2401 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     2461 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgtgtcacg
     2521 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     2581 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     2641 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     2701 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     2761 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     2821 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     2881 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     2941 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     3001 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     3061 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct tgatcccctg
     3121 cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc
     3181 ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag
     3241 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     3301 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     3361 ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag
     3421 cgtgaagct
//
