LOCUS       sequence-228321-        5974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre activity reporter plasmid; Zsgreen is expressed in cells without
            Cre, mCherry is expressed with Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5974)
  AUTHORS   Yang YS, Hughes TE
  TITLE     Cre stoplight: a red/green fluorescent reporter of Cre recombinase 
            expression in living cells.
  JOURNAL   Biotechniques. 2001 Nov;31(5):1036, 1038, 1040-1.
  PUBMED    11730010
REFERENCE   2  (bases 1 to 5974)
  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: 
            "Biotechniques."; date: "2001-11"; volume: "31(5)"; pages: "1036, 
            1038, 1040-1"
COMMENT     Sequence Label: Cre Stoplight 2.4
FEATURES             Location/Qualifiers
     source          1..5974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..209
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     160..180
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     254..273
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        254..272
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(292..325)
                     /label=loxH
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021). The loxH variant 
                     was optimized for expression constructs but remains 
                     compatible with loxP."
     CDS             339..1034
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    1047..1254
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1311..1329)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1325..1372
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    complement(1417..1450)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1470..2177
                     /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"
     CDS             2203..2910
                     /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"
     polyA_signal    3038..3159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3075..3094)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3129..3148
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3208..3224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3208..3224)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3221..3243)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3232..3248
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3256..3262,3263..3280,3281..3286))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3301..3322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3439..3456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3610..4198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3690..3709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4369..5160,5161..5229))
                     /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     4992..5011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5230..5334)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5409..5428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5600..5974,1..5)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
       61 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      121 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      181 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      241 ggcttatcga aattaatacg actcactata gggagaccca agctggctag aattacctca
      301 tatagcatac attatacgaa gttatgggcg cgctcgagat ggcccagtcc aagcacggcc
      361 tgaccaagga gatgaccatg aagtaccgca tggagggctg cgtggacggc cacaagttcg
      421 tgatcaccgg cgagggcatc ggctacccct tcaagggcaa gcaggccatc aacctgtgcg
      481 tggtggaggg cggccccttg cccttcgccg aggacatctt gtccgccgcc ttcatgtacg
      541 gcaaccgcgt gttcaccgag tacccccagg acatcgtcga ctacttcaag aactcctgcc
      601 ccgccggcta cacctgggac cgctccttcc tgttcgagga cggcgccgtg tgcatctgca
      661 acgccgacat caccgtgagc gtggaggaga actgcatgta ccacgagtcc aagttctacg
      721 gcgtgaactt ccccgccgac ggccccgtga tgaagaagat gaccgacaac tgggagccct
      781 cctgcgagaa gatcatcccc gtgcccaagc agggcatctt gaagggcgac gtgagcatgt
      841 acctgctgct gaaggacggt ggccgcttgc gctgccagtt cgacaccgtg tacaaggcca
      901 agtccgtgcc ccgcaagatg cccgactggc acttcatcca gcacaagctg acccgcgagg
      961 accgcagcga cgccaagaac cagaagtggc acctgaccga gcacgccatc gcctccggct
     1021 ccgccttgcc ctgactcgag cgcgccctgt gccttctagt tgccagccat ctgttgtttg
     1081 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1141 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1201 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatctaga
     1261 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1321 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1381 aggaactata tccggatatc cagcggccgc ggtaccataa cttcgtatag catacattat
     1441 acgaagttat gtttcggaat tcgcgcgcaa tggtgagcaa gggcgaggag gataacatgg
     1501 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1561 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1621 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     1681 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     1741 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1801 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1861 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1921 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     1981 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     2041 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     2101 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2161 tggacgagct gtacaagggg cgcgcctagg aattcgcgcg caatggtgag caagggcgag
     2221 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2281 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2341 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2401 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     2461 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2521 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     2581 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     2641 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     2701 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     2761 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     2821 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     2881 tccaccggcg gcatggacga gctgtacaag gggcgcgcct aggaattcga ttccacccgc
     2941 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3001 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     3061 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3121 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     3181 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3241 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3301 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3361 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3421 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3481 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3541 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4021 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4141 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4201 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4261 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4321 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4381 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4441 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4501 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4561 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4621 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4681 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4741 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4801 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4861 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4921 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4981 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5041 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5101 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5161 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5221 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5281 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5341 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     5401 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     5461 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     5521 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     5581 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     5641 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5701 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5761 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5821 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5881 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5941 tggcagtaca tctacgtatt agtcatcgct atta
//
