LOCUS       sequence-77875-e        5599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1-responsive fluorescent reporter plasmid containing reverse 
            complementary ZsGreen flanked by Bxb1 attB/P recognition sites in 
            head-to-head orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  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 5599)
  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-attBr-ZsGreen-attP
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..673
                     /label=chicken beta-actin promoter
     intron          674..1682
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    1695..1744
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     CDS             complement(1752..2447)
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTNNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     protein_bind    2460..2507
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     primer_bind     complement(2527..2544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2533..2757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2829..2846)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2829..2845)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2838..2860)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2987..3442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3074..3093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3284..3305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3468..3572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3573..3641,3642..4433)
                     /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(3791..3810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4604..5192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5093..5112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5346..5363
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5480..5501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5516..5521,5522..5539,5540..5546)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5554..5570
                     /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     5559..5581
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5578..5594
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5578..5594
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccgg attcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
      421 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
      481 attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg
      541 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc gcgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
      841 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
      901 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg cgaggggagc
     1021 gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga ggggaacaaa ggctgcgtgc
     1081 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc ggcggtcggg ctgtaacccc
     1141 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1201 gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1261 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctggcggag ccgaaatctg ggaggcgccg
     1441 ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccatctcca gcctcggggc
     1561 tgccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1681 agatccttaa ttaatcggcc ggcttgtcga cgacggcggt ctccgtcgtc aggatcatcc
     1741 gggctgacgg ttcagggcaa ggcggagccg gaggcgatgg cgtgctcggt caggtgccac
     1801 ttctggttct tggcgtcgct gcggtcctcg cgggtcagct tgtgctggat gaagtgccag
     1861 tcgggcatct tgcggggcac ggacttggcc ttgtacacgg tgtcgaactg gcagcgcaag
     1921 cggccaccgt ccttcagcag caggtacatg ctcacgtcgc ccttcaagat gccctgcttg
     1981 ggcacgggga tgatcttctc gcaggagggc tcccagttgt tggtcatctt cttcatcacg
     2041 gggccgtcgg cggggaagtt cacgccgtag aacttggact cgtggtacat gcagttctcc
     2101 tccacgctca cggtgatgtc ggcgttgcag atgcacacgg cgccgtcctc gaacaggaag
     2161 gagcggtccc aggtgtagcc ggcggggcag gagttcttga agtagtcgac gatgtcctgg
     2221 gggtactcgg tgaacacgcg gttgccgtac atgaaggcgg cggacaagat gtcctcggcg
     2281 aagggcaagg ggccgccctc caccacgcac aggttgatgg cctgcttgcc cttgaagggg
     2341 tagccgatgc cctcgccggt gatcacgaac ttgtggccgt ccacgcagcc ctccatgcgg
     2401 tacttcatgg tcatctcctt ggtcaggccg tgcttggact gggccatggt ggtggcggcg
     2461 gtttgtaccg tacaccactg agaccgcggt ggttgaccag acaaaccacg agagctcgct
     2521 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2581 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2641 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2701 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2761 ctgaggcgga aagaaccagc tggggctcga gatccactag ttctagcctc gaggctagag
     2821 cggccgccac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2881 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2941 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3001 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3061 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3181 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3301 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3361 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3421 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3481 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3541 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3601 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3661 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3721 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3781 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3841 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3901 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3961 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4021 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4081 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4141 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4201 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4261 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4321 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4381 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4441 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4501 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4561 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4621 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4681 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4741 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4801 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4861 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4921 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4981 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5041 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5101 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5161 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5221 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5281 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5341 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5401 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5461 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5521 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5581 gaaacagcta tgaccatga
//
