LOCUS       sequence-334132-        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             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"
     rep_origin      complement(1801..2389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1881..1900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2560..3351,3352..3420))
                     /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     3183..3202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3421..3525)
                     /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 cggccctatg gccaagttga ccagtgccgt tccggtgctc
     1261 accgcgagag acgtcgccgg agcggtcgag ttctggaccg acagactcgg gttctccaga
     1321 gacttcgtgg aggacgactt cgccggtgtg gtcagagacg acgtgaccct gttcatcagc
     1381 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gagaggcctg
     1441 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttcagaga cgcctccgga
     1501 ccggccatga ccgagatcgg cgagcagccg tggggaagag agttcgccct gagagacccg
     1561 gccggcaact gcgtgcactt cgtggccgag gagcaggact aatgctcgct ttcttgctgt
     1621 ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg ggggatatta
     1681 tgaagggcct tgagcatctg gattctgcct aataaaaaac atttattttc attgcaaaac
     1741 catgggagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3541 agtgccacct gacgtc
//
