LOCUS       sequence-337177-        6047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre under Prok2 minimal promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6047)
  TITLE     Prok2 Cre
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6047)
  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            "Unpublished"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: pProk2.Cre.T2A.mCherry
FEATURES             Location/Qualifiers
     source          1..6047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..113)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    102..309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   317..457
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   317..446
                     /label=AAV2 ITR
     rep_origin      532..987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(619..638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     829..850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1004..1023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1123..1145
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1183..1201)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1269..1373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1374..1442,1443..2234)
                     /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(1592..1611)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2405..2993
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2894..2913
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3055..3195
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3066..3195
                     /label=AAV2 ITR
     protein_bind    3202..3222
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3633..3657
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3658..3667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3670..3690
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3691..4716
                     /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             4726..4779
                     /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             4780..5490
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4925..4943)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5193..5212
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(5491..5515)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(5525..6047,1..66)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5578..5598)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5996..6007)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
       61 ccccgcatcg ggaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
      121 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
      181 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
      241 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
      301 tgctggggag ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
      361 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
      421 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
      481 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
      541 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      601 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      661 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      721 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
      781 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      841 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
      901 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      961 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     1021 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     1081 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1141 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     1201 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1261 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1321 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1381 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1441 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1501 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1561 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1621 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1681 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1741 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1801 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1861 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1921 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1981 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2041 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     2101 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2161 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2221 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2281 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2341 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2401 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2461 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2521 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2581 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2641 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2701 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2761 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2821 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2881 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2941 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3001 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca
     3061 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     3121 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     3181 tcactagggg ttccttctag acaactttgt atagaaaagt tgggcacgtc gtgggtaccc
     3241 cctgcacatc tgtatggcaa atgatgtgcc tctgcgagtg tgggggctga gcacgtgagg
     3301 ctctggaaaa caggacggcg aaggaggagg gtttctgaga ccacaaaagc ttcaggaagg
     3361 ctggctaggg ctgcggcgcc ccgcgggggc tctgcccgcg tggcgctttg cgcgtggggc
     3421 gcggggcacg tgcgcgtgtg cgcgtggagc gcggggtgtg tgcccgcgcc gtgccccccg
     3481 cgtgctgcct ggcgtgagtc accgcggggc tcgcctttat aaccgccgcc aggctcgcag
     3541 ctccgcagag cagcccggcc acgcgtcccc aacgtcccgg gtcccaacgc cccggaacgc
     3601 gtcccctaac cgccaccgcg tccccgggac gcccaagttt gtacaaaaaa gcaggctgcc
     3661 accatgggcc caaagaagaa gagaaaggtt tcgaatttac tgaccgtaca ccaaaatttg
     3721 cctgcattac cggtcgatgc aacgagtgat gaggttcgca agaacctgat ggacatgttc
     3781 agggatcgcc aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg
     3841 tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt
     3901 cgcgattatc ttctatatct tcaggcgcgc ggtctggcag taaaaactat ccagcaacat
     3961 ttgggccagc taaacatgct tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat
     4021 gctgtttcac tggttatgcg gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca
     4081 aaacaggctc tagcgttcga acgcactgat ttcgaccagg ttcgttcact catggaaaat
     4141 agcgatcgct gccaggatat acgtaatctg gcatttctgg ggattgctta taacaccctg
     4201 ttacgtatag ccgaaattgc caggatcagg gttaaagata tctcacgtac tgacggtggg
     4261 agaatgttaa tccatattgg cagaacgaaa acgctggtta gcaccgcagg tgtagagaag
     4321 gcacttagcc tgggggtaac taaactggtc gagcgatgga tttccgtctc tggtgtagct
     4381 gatgatccga ataactacct gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct
     4441 gccaccagcc agctatcaac tcgcgccctg gaagggattt ttgaagcaac tcatcgattg
     4501 atttacggcg ctaaggatga ctctggtcag agatacctgg cctggtctgg acacagtgcc
     4561 cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt caataccgga gatcatgcaa
     4621 gctggtggct ggaccaatgt aaatattgtc atgaactata tccgtaacct ggatagtgaa
     4681 acaggggcaa tggtgcgcct gctggaagat ggcgatggaa gcggagaggg caggggaagt
     4741 cttctaacat gcggggacgt ggaggaaaat cccggcccca tggtgagcaa gggcgaggag
     4801 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     4861 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4921 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     4981 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     5041 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     5101 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     5161 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     5221 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     5281 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     5341 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     5401 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     5461 accggcggca tggacgagct gtacaagtaa acccagcttt cttgtacaaa gtgggaattc
     5521 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5581 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5641 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5701 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5761 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5821 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5881 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5941 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6001 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgc
//
