LOCUS       sequence-340324-        3556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of CRE-T2A-BleoR synthetic construct for the 
            excision of floxed cassettes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3556)
  TITLE     Verstreken lab plasmids
REFERENCE   2  (bases 1 to 3556)
  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-BleoR
FEATURES             Location/Qualifiers
     source          1..3556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             122..142
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             143..1168
                     /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             1178..1231
                     /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"
     rep_origin      complement(1805..2393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1885..1904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2564..3355,3356..3424))
                     /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     3187..3206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3425..3529)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtctaatac gactcactat aaggacattt gcttctgaca caactgtgtt cactagcaac
       61 ctcaaacaga attcgagctc ggtacccggg gatcctctag agtcaaagaa ttgccgccat
      121 gcccaagaag aagaggaagg tgtccaattt actgaccgta caccaaaatt tgcctgcatt
      181 accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg
      241 ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc
      301 atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta
      361 tcttctatat cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca
      421 gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc
      481 actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc
      541 tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg
      601 ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat
      661 agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt
      721 aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag
      781 cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc
      841 gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag
      901 ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg
      961 cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg
     1021 agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg
     1081 ctggaccaat gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc
     1141 aatggtgcgc ctgctggaag atggcgatgg cagcggagag ggcagaggaa gtcttctaac
     1201 atgcggtgac gtggaggaga atcccggccc tatggccaag ttgaccagtg ccgttccggt
     1261 gctcaccgcg agagacgtcg ccggagcggt cgagttctgg accgacagac tcgggttctc
     1321 cagagacttc gtggaggacg acttcgccgg tgtggtcaga gacgacgtga ccctgttcat
     1381 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgagagg
     1441 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttca gagacgcctc
     1501 cggaccggcc atgaccgaga tcggcgagca gccgtgggga agagagttcg ccctgagaga
     1561 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactaatgct cgctttcttg
     1621 ctgtccaatt tctattaaag gttcctttgt tccctaagtc caactactaa actgggggat
     1681 attatgaagg gccttgagca tctggattct gcctaataaa aaacatttat tttcattgca
     1741 aaaccatggg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2221 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2401 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2461 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2521 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2581 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2641 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2701 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2761 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2821 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2881 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2941 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3001 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3061 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3121 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3181 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3241 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3301 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3361 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3421 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3481 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3541 gaaaagtgcc acctga
//
