LOCUS       sequence-14380-e        2997 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 2997)
  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 2997)
  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-Blast
FEATURES             Location/Qualifiers
     source          1..2997
                     /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    536..657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(547..566)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     601..620
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(815..1213)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(1232..1279)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1327..1656)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1341..1476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1395..1414)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1641..1661
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(1848..2642)
                     /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"
     primer_bind     complement(1959..1978)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2569..2588
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
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 gcggccgcaa ttgttaacag atccgtcgac ggtatacaga catgataaga
      541 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      601 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttggg
      661 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
      721 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtag
      781 cacgtgtcag tcctgctcct cggccacgaa gtgcttagcc ctcccacaca taaccagagg
      841 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
      901 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
      961 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     1021 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     1081 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     1141 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     1201 aaggcttggc catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
     1261 ccgatgatta attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg
     1321 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     1381 agaggcagag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     1441 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     1501 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     1561 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     1621 ggagcctggg gactttccac accctaactg acacacattc cacagaatta tcccggggtg
     1681 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     1741 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     1801 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     1861 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     1921 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     1981 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     2041 ccattttcca ccatgatatt cggcaagcag gcatcgccat gtgtcacgac gagatcctcg
     2101 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     2161 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     2221 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     2281 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     2341 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     2401 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     2461 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     2521 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     2581 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     2641 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
     2701 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     2761 gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc
     2821 catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag
     2881 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     2941 gtgttccgct tcctttagca gcccttgcgc cctgagtgct tgcggcagcg tgaagct
//
