LOCUS       sequence-491856-        6507 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Cre-responsive fluorescent reporter plasmid containing a 
            STOP-cassette flanked by loxP sites followed by ZsGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6507)
  AUTHORS   Hermann M, Stillhard P, Wildner H, Seruggia D, Kapp V, 
            Sanchez-Iranzo H, Mercader N, Montoliu L, Zeilhofer HU, Pelczar P
  TITLE     Binary recombinase systems for high-resolution conditional 
            mutagenesis.
  JOURNAL   Nucleic Acids Res. 2014 Jan 9.
  PUBMED    24413561
REFERENCE   2  (bases 1 to 6507)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2014 Jan 9."
COMMENT     Sequence Label: pCAG-loxPSTOPloxP-ZsGreen
FEATURES             Location/Qualifiers
     source          1..6507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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)."
     primer_bind     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        99..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        480..756
                     /label=chicken beta-actin promoter
     intron          757..1773
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1697..1719
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1786..1805
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1786..1804
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1833..1866
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2033..2167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2287..2421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2541..2675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2704..2737
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2798..2807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2804..3499
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     primer_bind     complement(3519..3536)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3525..3749
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3821..3838)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3821..3837)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3830..3852)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3978..4433)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4115..4136)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4327..4346
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4460..4564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4565..4633,4634..5425)
                     /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(4783..4802)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5596..6184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6085..6104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6338..6355
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6472..6493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgaggcgcg ccggattcga cattgattat tgactagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
      541 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg
      601 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      661 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      721 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      781 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      841 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      901 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      961 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
     1021 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1081 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1141 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1201 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1261 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1321 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     1381 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1441 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1501 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1561 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1621 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1681 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1741 aaccatgttc atgccttctt ctttttccta cagatcctta attaataata cgactcacta
     1801 tagggggtcg acccgccacc aatcatttaa atataacttc gtataatgta tgctatacga
     1861 agttattcgc gatgaataaa tgaaagcttg cagatctgcg actctagagg atctgcgact
     1921 ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1981 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2161 ctggatctgc gactctagag gatcataatc agccatacca catttgtaga ggttttactt
     2221 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2281 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2341 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2401 gtatcttatc atgtctggat ctgcgactct agaggatcat aatcagccat accacatttg
     2461 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2521 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2581 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2641 ccaaactcat caatgtatct tatcatgtct ggatccccat caagctgatc cggaaccctt
     2701 aatataactt cgtataatgt atgctatacg aagttattag gtccctcgac ctgcagccca
     2761 agctagatcg aattcggccg gccgatatcg gcgcgccgcc accatggccc agtccaagca
     2821 cggcctgacc aaggagatga ccatgaagta ccgcatggag ggctgcgtgg acggccacaa
     2881 gttcgtgatc accggcgagg gcatcggcta ccccttcaag ggcaagcagg ccatcaacct
     2941 gtgcgtggtg gagggcggcc ccttgccctt cgccgaggac atcttgtccg ccgccttcat
     3001 gtacggcaac cgcgtgttca ccgagtaccc ccaggacatc gtcgactact tcaagaactc
     3061 ctgccccgcc ggctacacct gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat
     3121 ctgcaacgcc gacatcaccg tgagcgtgga ggagaactgc atgtaccacg agtccaagtt
     3181 ctacggcgtg aacttccccg ccgacggccc cgtgatgaag aagatgaccg acaactggga
     3241 gccctcctgc gagaagatca tccccgtgcc caagcagggc atcttgaagg gcgacgtgag
     3301 catgtacctg ctgctgaagg acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa
     3361 ggccaagtcc gtgccccgca agatgcccga ctggcacttc atccagcaca agctgacccg
     3421 cgaggaccgc agcgacgcca agaaccagaa gtggcacctg accgagcacg ccatcgcctc
     3481 cggctccgcc ttgccctgag tcagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3541 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3601 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3661 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3721 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3781 gagatccact agttctagcc tcgaggctag agcggccgcc actggccgtc gttttacaac
     3841 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3901 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3961 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4021 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4081 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4141 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4201 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4261 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4321 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4381 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4441 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4501 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4561 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4621 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4681 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4741 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4801 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4861 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4921 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4981 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5041 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5101 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5161 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5221 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5281 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5341 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5401 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5461 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5521 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5701 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5881 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5941 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6001 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6061 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6121 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6181 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6241 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6301 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6361 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6421 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6481 ttagctcact cattaggcac cccaggc
//
