LOCUS       sequence-337175-        7124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre under Prok receptor 2 minimal promoter in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7124)
  TITLE     Prok2 Cre
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7124)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pProkR2.Cre.T2A.Venus
FEATURES             Location/Qualifiers
     source          1..7124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..20)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9..216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   224..364
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   224..353
                     /label=AAV2 ITR
     rep_origin      439..894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(526..545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     736..757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(911..930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1030..1052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1090..1108)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1176..1280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1281..1349,1350..2141)
                     /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     complement(1499..1518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2312..2900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2801..2820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2962..3102
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2973..3102
                     /label=AAV2 ITR
     protein_bind    3109..3129
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    4608..4632
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4633..4642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4645..4665
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4666..5691
                     /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             5701..5754
                     /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             join(5755..5757,5758..5760,5761..6474)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6061..6080)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6408..6429
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6475..6499)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6509..7097
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6562..6582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6980..6991)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
       61 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
      121 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
      181 ggaggattgg gaagagaata gcaggcatgc tggggagggc cgcaggaacc cctagtgatg
      241 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
      301 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
      361 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      421 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      481 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
      541 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
      601 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
      661 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
      721 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
      781 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
      841 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
      901 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
      961 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     1021 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     1081 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1141 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1201 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1261 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1321 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1381 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1441 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1501 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1561 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1621 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1681 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1741 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1801 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1861 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1921 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1981 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2041 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2101 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2161 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2221 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2281 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2341 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2401 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2461 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2521 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2581 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2641 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2701 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2761 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2821 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2881 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2941 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg
     3001 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     3061 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttctagaca actttgtata
     3121 gaaaagttgc tgcacactta gtggaagcca caaatgcttg cagaggacag tgctccgtct
     3181 caaggatggc cagctgctga ggtcacaacg ggttcaagag cttacaaaac ccttggaagg
     3241 accacccgga agtctgagac cctaattcag tagatctagg aggcaggcgg gtctcggggg
     3301 gcggagtcgg gcgccgggag gggcgggaca cctgggctgg gtccggggct gttggatgag
     3361 ctgcagagcg aagcagcaac tccagaaaga ctgatgcgag gagagcagga ccaagcgaca
     3421 cacgcccacc aagtaggcaa gcctcaacca gagctagtgt tcccagagcc cgggccgcgc
     3481 ccacctatgg tggcgccacc ggccagggag tgcgtccctg aaagcaccga ggcaagtgag
     3541 taccgggttg cggcgggcgg gcggacttgg gggcactccg cgtctgttcc aggagatcac
     3601 ctcctacctc cgaaccctgg gattggggac taggggactg gagtggagcc ccgaaatcgc
     3661 ggactcagtg gaaggcgtac ggacaagggt gggggtatct gcgtgtttcc tgcgccctgc
     3721 aggcacctct ggtgcccggc tttgtttctt gagcttaaat tccccactgg gtacctaatc
     3781 ctgtaccagt gttgcactga gtgagtgaga gacgtgggaa ccttcaaacg ggctccctct
     3841 tcccacagtc tacgggctgg tgatgagaag gcggctggaa ctcccgctta ttggtccccg
     3901 gtggcgatct ttgggagacc aatagacgcc ccagagggag gacactggga tccagactcc
     3961 actggaaccc cgcttttcag gtagggtgga gcgtgcggct gattcctgtg tagcaaatcg
     4021 gggtttcact aacgccggca gagtggcgat tcctgtcccc cagcgctctc cctcgtaaaa
     4081 ctgggcagac ctcgctatcc agtttgatag cttcttgcta cccttacccg ttgctcccgt
     4141 agctcccgca ctggctggtg tgctgaggtg ggagaggaag gcagttcatt gagacagact
     4201 aagtcaggct ggagtcctct ctccagcccc tcgcttgccc catagcagct acctcctggg
     4261 agctagcatt ttggagactc tattttgtct cccttccggc taagggtgca tactcttctt
     4321 gggtttctct gaagattcca gttcattctg tgaccagcaa gcagacggcc tcccgtcttc
     4381 ttggccctgg gtgactctgg ttggactgta cctagctaac tcttgggggg actagccaga
     4441 tctgatgttt gctgaccagt ggttttatct gcccccagat cctggatggt atctgttctc
     4501 cctaaggatt ctaacaggga cctgcactca ctgaccccag cagaactgct gaactccacg
     4561 tgagcgcatc tccctgatac acaccagccc acctgtagca tcatcaacca agtttgtaca
     4621 aaaaagcagg ctgccaccat gggcccaaag aagaagagaa aggtttcgaa tttactgacc
     4681 gtacaccaaa atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac
     4741 ctgatggaca tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg
     4801 tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca
     4861 gaacctgaag atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa
     4921 actatccagc aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga
     4981 ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat
     5041 gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt
     5101 tcactcatgg aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt
     5161 gcttataaca ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca
     5221 cgtactgacg gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc
     5281 gcaggtgtag agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc
     5341 gtctctggtg tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt
     5401 gttgccgcgc catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa
     5461 gcaactcatc gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg
     5521 tctggacaca gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata
     5581 ccggagatca tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa ctatatccgt
     5641 aacctggata gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga tggaagcgga
     5701 gagggcaggg gaagtcttct aacatgcggg gacgtggagg aaaatcccgg ccccatggtg
     5761 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     5821 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     5881 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     5941 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     6001 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     6061 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     6121 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     6181 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     6241 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     6301 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     6361 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     6421 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaacccag
     6481 ctttcttgta caaagtggga attccgataa tcaacctctg gattacaaaa tttgtgaaag
     6541 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     6601 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6661 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6721 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6781 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6841 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6901 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6961 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     7021 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     7081 ttgggccgcc tccccgcatc gggaattcct agagctcgct gatc
//
