LOCUS       sequence-77877-e        6432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  B3-responsive fluorescent reporter plasmid containing a 
            STOP-cassette flanked by B3-recognition sites followed by ZsGreen 
            CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6432)
  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 6432)
  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: "Nucleic 
            Acids Res. 2014 Jan 9."
COMMENT     Sequence Label: pCAG-b3STOPb3-ZsGreen
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..33,34..40)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    48..64
                     /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     53..75
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     72..88
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     72..88
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        108..487
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        489..766
                     /label=chicken beta-actin promoter
     intron          767..1775
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1699..1721
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1788..1807
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1788..1806
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2022..2156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2276..2410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2530..2664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2733..2742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2739..3434
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     primer_bind     complement(3453..3470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3459..3683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3755..3772)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3755..3771)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3786)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3913..4368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4000..4019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4210..4231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4394..4498
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4499..4567,4568..5359)
                     /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(4717..4736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5530..6118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6019..6038
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6272..6289
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6406..6427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
       61 acaatttcac acaggaaaca gctatgacca tgaggcgcgc cggattcgac attgattatt
      121 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      181 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      241 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      301 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      361 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      421 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      481 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
      541 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
      601 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
      661 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      721 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgtt
      781 gccttcgccc cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac
      841 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      901 cttggtttaa tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg
      961 ggagggccct ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
     1021 ggagcgccgc gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
     1081 gctttgtgcg ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     1141 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     1201 ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct
     1261 gagcacggcc cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg gctcgccgtg
     1321 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
     1381 gagggctcgg gggaggggcg cggcggcccc ggagcgccgg cggctgtcga ggcgcggcga
     1441 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1501 aaatctggcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcgggcg
     1561 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1621 cgccgtcccc ttctccatct ccagcctcgg ggctgccgca gggggacggc tgccttcggg
     1681 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     1741 ctaaccatgt tcatgccttc ttctttttcc tacagatcct taattaataa tacgactcac
     1801 tatagggggt cgacccgcca ccggttgctt aagaattact tattcttaag caacctcgcg
     1861 atgaataaat gaaagcttgc agatctgcga ctctagagga tctgcgactc tagaggatca
     1921 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1981 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2041 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2101 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgcg
     2161 actctagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2221 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2281 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2341 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2401 tgtctggatc tgcgactcta gaggatcata atcagccata ccacatttgt agaggtttta
     2461 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2521 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2581 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2641 aatgtatctt atcatgtctg gatccccatc aagctgatcc ggaaccctta atggttgctt
     2701 aagaattact tattcttaag caaccggcag ccgccaccat ggcccagtcc aagcacggcc
     2761 tgaccaagga gatgaccatg aagtaccgca tggagggctg cgtggacggc cacaagttcg
     2821 tgatcaccgg cgagggcatc ggctacccct tcaagggcaa gcaggccatc aacctgtgcg
     2881 tggtggaggg cggccccttg cccttcgccg aggacatctt gtccgccgcc ttcatgtacg
     2941 gcaaccgcgt gttcaccgag tacccccagg acatcgtcga ctacttcaag aactcctgcc
     3001 ccgccggcta cacctgggac cgctccttcc tgttcgagga cggcgccgtg tgcatctgca
     3061 acgccgacat caccgtgagc gtggaggaga actgcatgta ccacgagtcc aagttctacg
     3121 gcgtgaactt ccccgccgac ggccccgtga tgaagaagat gaccgacaac tgggagccct
     3181 cctgcgagaa gatcatcccc gtgcccaagc agggcatctt gaagggcgac gtgagcatgt
     3241 acctgctgct gaaggacggt ggccgcttgc gctgccagtt cgacaccgtg tacaaggcca
     3301 agtccgtgcc ccgcaagatg cccgactggc acttcatcca gcacaagctg acccgcgagg
     3361 accgcagcga cgccaagaac cagaagtggc acctgaccga gcacgccatc gcctccggct
     3421 ccgccttgcc ctgaaccgag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     3481 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3541 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3601 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3661 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgagatc
     3721 cactagttct agcctcgagg ctagagcggc cgccactggc cgtcgtttta caacgtcgtg
     3781 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3841 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3901 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3961 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4081 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4141 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4201 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4261 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4321 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     4381 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4441 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4501 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4561 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4621 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4681 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4741 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4801 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4861 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4921 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4981 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5041 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5101 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5161 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5221 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5281 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5341 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5401 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5461 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5521 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5581 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5641 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5701 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5761 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5821 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5881 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5941 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6001 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6061 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6121 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6181 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6241 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6301 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6361 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6421 cactcattag gc
//
