LOCUS       sequence-419563-        5864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cre and mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5864)
  TITLE     Vector expressing Cre and mCherry
REFERENCE   2  (bases 1 to 5864)
  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: pCre-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..5864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     283..302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        557..860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        861..1064
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1015..1035
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1166..2194
                     /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="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2252..2308
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2315..3025
                     /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(2460..2478)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2728..2747
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3148..3269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3185..3204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3239..3258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3276..3731)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3413..3434)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3625..3644
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3758..3862
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3864..4221
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3864..4055
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3887..3907)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4072..4207
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4256..5050
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4256..5050
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4310..4329)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4920..4939
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5238..5257)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5282..5329
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5658..5864,1..382)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
       61 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      421 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      481 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      541 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      601 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      661 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      721 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      781 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      841 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      901 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      961 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     1021 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     1081 agatccgcta gcgctaccgg actcagatct cgaggggcag agccgatcct gtacacttta
     1141 cttaaaacca ttatctgagt gtgaaatgtc caatttactg accgtacacc aaaatttgcc
     1201 tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag aacctgatgg acatgttcag
     1261 ggatcgccag gcgttttctg agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg
     1321 ggcggcatgg tgcaagttga ataaccggaa atggtttccc gcagaacctg aagatgttcg
     1381 cgattatctt ctatatcttc aggcgcgcgg tctggcagta aaaactatcc agcaacattt
     1441 gggccagcta aacatgcttc atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc
     1501 tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa
     1561 acaggctcta gcgttcgaac gcactgattt cgaccaggtt cgttcactca tggaaaatag
     1621 cgatcgctgc caggatatac gtaatctggc atttctgggg attgcttata acaccctgtt
     1681 acgtatagcc gaaattgcca ggatcagggt taaagatatc tcacgtactg acggtgggag
     1741 aatgttaatc catattggca gaacgaaaac gctggttagc accgcaggtg tagagaaggc
     1801 acttagcctg ggggtaacta aactggtcga gcgatggatt tccgtctctg gtgtagctga
     1861 tgatccgaat aactacctgt tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc
     1921 caccagccag ctatcaactc gcgccctgga agggattttt gaagcaactc atcgattgat
     1981 ttacggcgct aaggatgact ctggtcagag atacctggcc tggtctggac acagtgcccg
     2041 tgtcggagcc gcgcgagata tggcccgcgc tggagtttca ataccggaga tcatgcaagc
     2101 tggtggctgg accaatgtaa atattgtcat gaactatatc cgtaacctgg atagtgaaac
     2161 aggggcaatg gtgcgcctgc tggaagatgg cgatgtcgaa attctgcagt cgacggtacc
     2221 gcgggcccgg gatccaccgg tgggatcggg tgctactaac ttcagcctgc tgaagcaggc
     2281 tggagacgtg gaggagaacc ctggacctgg gtccatggtg agcaagggcg aggaggataa
     2341 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     2401 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2461 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2521 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2581 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2641 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2701 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2761 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2821 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     2881 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     2941 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     3001 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     3061 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3121 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3241 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3301 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3361 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3421 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3481 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3541 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3601 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3661 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3721 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3781 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3841 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3901 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3961 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4021 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4081 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4141 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4201 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4261 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4321 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4381 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4441 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4501 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4561 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4621 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4681 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4741 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4801 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4861 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4921 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4981 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5041 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5101 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5161 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5221 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5281 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5341 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5401 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5461 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5521 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5581 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5641 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5701 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5761 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5821 tagttaggcc accacttcaa gaactctgta gcaccgccta cata
//
