LOCUS       sequence-334130-        3813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of CRE-T2A-PuroR synthetic construct for the 
            excision of floxed cassettes .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3813)
  TITLE     Verstreken lab plasmids
REFERENCE   2  (bases 1 to 3813)
  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"
COMMENT     Sequence Label: pIVTR(T7A)-CRE-T2A-PuroR
FEATURES             Location/Qualifiers
     source          1..3813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             118..138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             139..1164
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             1174..1227
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1237..1836
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1237..1256)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1733..1753
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     rep_origin      complement(2058..2646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2138..2157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2817..3608,3609..3677))
                     /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     3440..3459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3678..3782)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taatacgact cactataagg acatttgctt ctgacacaac tgtgttcact agcaacctca
       61 aacagaattc gagctcggta cccggggatc ctctagagtc aaagaattgc cgccatgccc
      121 aagaagaaga ggaaggtgtc caatttactg accgtacacc aaaatttgcc tgcattaccg
      181 gtcgatgcaa cgagtgatga ggttcgcaag aacctgatgg acatgttcag ggatcgccag
      241 gcgttttctg agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg
      301 tgcaagttga ataaccggaa atggtttccc gcagaacctg aagatgttcg cgattatctt
      361 ctatatcttc aggcgcgcgg tctggcagta aaaactatcc agcaacattt gggccagcta
      421 aacatgcttc atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc tgtttcactg
      481 gttatgcggc ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta
      541 gcgttcgaac gcactgattt cgaccaggtt cgttcactca tggaaaatag cgatcgctgc
      601 caggatatac gtaatctggc atttctgggg attgcttata acaccctgtt acgtatagcc
      661 gaaattgcca ggatcagggt taaagatatc tcacgtactg acggtgggag aatgttaatc
      721 catattggca gaacgaaaac gctggttagc accgcaggtg tagagaaggc acttagcctg
      781 ggggtaacta aactggtcga gcgatggatt tccgtctctg gtgtagctga tgatccgaat
      841 aactacctgt tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc caccagccag
      901 ctatcaactc gcgccctgga agggattttt gaagcaactc atcgattgat ttacggcgct
      961 aaggatgact ctggtcagag atacctggcc tggtctggac acagtgcccg tgtcggagcc
     1021 gcgcgagata tggcccgcgc tggagtttca ataccggaga tcatgcaagc tggtggctgg
     1081 accaatgtaa atattgtcat gaactatatc cgtaacctgg atagtgaaac aggggcaatg
     1141 gtgcgcctgc tggaagatgg cgatggcagc ggagagggca gaggaagtct tctaacatgc
     1201 ggtgacgtgg aggagaatcc cggccctagg ctcgagatga ccgagtacaa gcccacggtg
     1261 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     1321 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     1381 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     1441 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     1501 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     1561 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     1621 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     1681 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     1741 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     1801 cgcacctggt gcatgacccg caagcccggt gcctgatctg acctgcaggc atgcaagctt
     1861 gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac
     1921 taaactgggg gatattatga agggccttga gcatctggat tctgcctaat aaaaaacatt
     1981 tattttcatt gcaaaaccat gggagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2221 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2461 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2881 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2941 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3001 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3061 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3781 cgcacatttc cccgaaaagt gccacctgac gtc
//
