LOCUS       sequence-483038-        6508 bp    DNA     circular SYN 12-AUG-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 6508)
  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 6508)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 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..6508
                     /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"
     intron          758..1774
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1698..1720
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1787..1806
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1787..1805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1834..1867
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2034..2168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2288..2422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2542..2676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2705..2738
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2799..2808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2805..3500
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     primer_bind     complement(3520..3537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3526..3750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3822..3839)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3822..3838)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3831..3853)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3979..4434)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4116..4137)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4328..4347
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4461..4565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4566..4634,4635..5426)
                     /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(4784..4803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5597..6185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6086..6105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6339..6356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6473..6494
                     /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 gcgcgcgccg gggggggggg gggggggggg gggggggggg ggggsgrggs ggrgrggtgc
      661 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      721 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
      781 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      841 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
      901 ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg
      961 ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
     1021 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
     1081 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
     1141 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     1201 tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca
     1261 cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg
     1321 cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggrggg
     1381 ctcgggggrg gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg
     1441 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     1501 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
     1561 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     1621 ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg
     1681 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
     1741 taaccatgtt catgccttct tctttttcct acagatcctt aattaataat acgactcact
     1801 atagggggtc gacccgccac caatcattta aatataactt cgtataatgt atgctatacg
     1861 aagttattcg cgatgaataa atgaaagctt gcagatctgc gactctagag gatctgcgac
     1921 tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1981 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     2161 tctggatctg cgactctaga ggatcataat cagccatacc acatttgtag aggttttact
     2221 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2281 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2401 tgtatcttat catgtctgga tctgcgactc tagaggatca taatcagcca taccacattt
     2461 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2521 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2581 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2641 tccaaactca tcaatgtatc ttatcatgtc tggatcccca tcaagctgat ccggaaccct
     2701 taatataact tcgtataatg tatgctatac gaagttatta ggtccctcga cctgcagccc
     2761 aagctagatc gaattcggcc ggccgatatc ggcgcgccgc caccatggcc cagtccaagc
     2821 acggcctgac caaggagatg accatgaagt accgcatgga gggctgcgtg gacggccaca
     2881 agttcgtgat caccggcgag ggcatcggct accccttcaa gggcaagcag gccatcaacc
     2941 tgtgcgtggt ggagggcggc cccttgccct tcgccgagga catcttgtcc gccgccttca
     3001 tgtacggcaa ccgcgtgttc accgagtacc cccaggacat cgtcgactac ttcaagaact
     3061 cctgccccgc cggctacacc tgggaccgct ccttcctgtt cgaggacggc gccgtgtgca
     3121 tctgcaacgc cgacatcacc gtgagcgtgg aggagaactg catgtaccac gagtccaagt
     3181 tctacggcgt gaacttcccc gccgacggcc ccgtgatgaa gaagatgacc gacaactggg
     3241 agccctcctg cgagaagatc atccccgtgc ccaagcaggg catcttgaag ggcgacgtga
     3301 gcatgtacct gctgctgaag gacggtggcc gcttgcgctg ccagttcgac accgtgtaca
     3361 aggccaagtc cgtgccccgc aagatgcccg actggcactt catccagcac aagctgaccc
     3421 gcgaggaccg cagcgacgcc aagaaccaga agtggcacct gaccgagcac gccatcgcct
     3481 ccggctccgc cttgccctga gtcagagctc gctgatcagc ctcgactgtg ccttctagtt
     3541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3721 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3781 cgagatccac tagttctagc ctcgaggcta gagcggccgc cactggccgt cgttttacaa
     3841 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3901 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3961 agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4021 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4081 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4141 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4201 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4261 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4321 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4381 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4441 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4561 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5101 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5281 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5401 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6301 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6361 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6421 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     6481 gttagctcac tcattaggca ccccaggc
//
