LOCUS       sequence-121687-        7987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway destination vector for phiC31-mediated 
            transgenesis in zebrafish, harbors ubiquitin promoter (ubi)-driven 
            EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7987)
  AUTHORS   Mosimann C, Puller AC, Lawson KL, Tschopp P, Amsterdam A, Zon LI
  TITLE     Site-directed zebrafish transgenesis into single landing sites with 
            the phiC31 integrase system.
  JOURNAL   Dev Dyn. 2013 Aug;242(8):949-63. doi: 10.1002/dvdy.23989. Epub 2013 
            Jul 3.
  PUBMED    23723152
REFERENCE   2  (bases 1 to 7987)
  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"; doi: 
            "10.1002/dvdy.23989"; journalName: "Dev Dyn"; date: "2013-08"; 
            volume: "242"; issue: "8"; pages: "949-63"
COMMENT     Sequence Label: pDestattB_ubi:EGFP (pCM272)
FEATURES             Location/Qualifiers
     source          1..7987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(252..270)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(291..307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(304..326)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    315..331
                     /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(339..345,346..363,364..369))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    384..405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(522..539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(693..1281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(773..792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1452..2243,2244..2312))
                     /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     2075..2094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2313..2417)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2443..2898)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2580..2601)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2792..2811
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3025..3047
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3040..3056
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3063..3082
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3063..3081
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3115..3133)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     primer_bind     3241..3257
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3277..3297
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    6831..6855
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(6861..6863,6864..6866,6867..7577)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6861..6863,6864..6866,6867..7577)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6906..6927)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7167..7186)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7514..7535
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(7639..7663)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7665..7799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7689..7708)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7743..7762
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(7862..7882)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(7890..7906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(7951..7987,1..33)
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 ggctccccgg gcgcgtactc cacctcaccc atctggtcca tcatgatgaa cgggtcgagg
       61 tggcggtagt tgatcccggc gaacgcgcgg cgcaccggga agccctcgcc ctcgaaaccg
      121 ctgggcgcgg tggtcacggt gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg
      181 cggggcagcg tcagcgggtt ctcgacggtc acggcgggca tgtcgacact agttctagcc
      241 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
      301 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      361 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      421 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      481 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      541 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      601 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      661 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      721 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      781 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      841 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      901 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      961 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1021 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1081 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1141 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1201 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1261 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1321 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1381 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1441 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1501 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1561 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1621 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1681 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1741 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1801 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1861 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1921 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1981 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2041 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2101 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2161 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2221 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2281 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2341 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2401 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     2461 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2521 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2581 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2641 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2701 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2761 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2821 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2881 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2941 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     3001 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     3061 tgtaatacga ctcactatag ggcgaattgg gtacgtaccg ggcccctagt atgtatgtaa
     3121 gttaataaaa cccatttttg cggaaagtag ataaaaaaaa catttttttt ttttactgca
     3181 ctggatctag cggatccggg aattgggaat tcgttaacag atctgcggcc gcggctcgag
     3241 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttggct
     3301 ccgaattcgc ccttaaactc gagaccagca aagttctaga atttgtcgaa acatttatgt
     3361 tatatatttc ctgaaaaaaa ttctgagtaa gttcttaagt gttattgcca gcaacataaa
     3421 caacagacgg caaaatgaat aaatgataac aaagcagtag gcttaaataa acctaatttt
     3481 tataggctgt tctctacaac cctcaaacag tgattagttt tgtacttata aacttgccct
     3541 ttcattcata tttcaagaaa attggttcag aagatctgga tattctagca gttgttcaag
     3601 ctcatggagg gatcagtgac ctgattccac aatgactagg cctaatccag aaattagatg
     3661 actgtcaaca taaaaaggca cagcactcac tagctgccct atatatttta ttatatttta
     3721 catatattat tttatttatt tagctctgag tgctgtactt tctggttaaa gaaaactgct
     3781 tacaacagct aacctgtact acctcaggct cagggaattt ggaacaggtt tgtctggttt
     3841 gtttctttaa ccatgcatgc ttgttttcaa ctatggcaac acagtcacat gggacattac
     3901 agaaatgatt tgtcgatgac atgcgacttt tctttaataa agcgcaaaga tcccaaaaag
     3961 caaactttta acaaaaatca tataattata ttttcaatcc agctttgtag caactttgtg
     4021 ctgctgttca ctcagcaaca gatagtcagt ataaggtcag tgtgtctcaa agcagtgcca
     4081 tctgtttcac acattgcgtt ctatatataa gtgtgctggt tgacacgaca ctgtataagg
     4141 cctaggctaa aacacaaaca atgtagaatg acactgtgtt ttttttgtaa acaaatgttg
     4201 tttttggtta aacatctttg tgaaaacatc ctcctgtcat gtatttgcta tattcaaatg
     4261 ttaaacccgt gcagaataga acatatacaa aaaaaaacaa cacaacacat ttttaaacat
     4321 tattaaatat caagtattgc tggcagttct gtttctgttt tacagtaccc tttgccacag
     4381 ttctccgctt ttcctggtcc agattccaca agtctgattc accaatagca aagcgaataa
     4441 acaaccaaag cagccaatca ctgcttgtag actgtcctgc gagaccggcc cattccagca
     4501 cattctggaa acttccttta tatgataatt ataaatacat ttaaattatt gatacaaaac
     4561 atgtaattcc tagaacataa ccatagcaat cattagtttt cagggtaatt atgtattttt
     4621 aggatttgac tgcggaaaga tctggtcatg tgacgtctca tgaacgtcac ggccctgggt
     4681 ttctataaat acagtaggac tctcgaccat cggcagattt ttcgaagaag aagatcagtt
     4741 tcaggagccg tactgttccg tttcaacgca aatattaacg gtaagagcga atttcctagt
     4801 ttgttttcat gccattcttt aaaaccatag cgtattactt taattatagt aaactttcgc
     4861 tttctttatt acaagagacg ttttgtgttg attctccgcg gacattttcg gtcagacaat
     4921 cagaaaatga ccgcggagga ccagtaactt gcattacacg taagttaaat cttcgtgtat
     4981 taaaatggtt aggttgttaa cgtcaaatag gttaccgtgt ttgcgtgtga tcaggttggt
     5041 tttgttagat ttttgtcagt atttttaatt tatttgtttt agtttattta ttttttttgc
     5101 tgaatcatag tttgtgaaca aagaacccgg atgttacata cagtacagcc gccatgttac
     5161 agagagttat aacttaatca ttttaaaaat aattttgcct tacttttagt ttgtcatgtt
     5221 gagaaatgag gaaatgttaa aatgaggaaa tatccaatta atttaatata tcaaaataat
     5281 ccatgattac aatgcactga actggagaaa attaagatgt tttctagtgt catgaaacaa
     5341 atgtaagaga tgtacattgt agatgtttta tgtcaagaat tggctagttg atgcagcata
     5401 ctggcgatac tcagttgtaa taacagtaac gttacatgtt aatagactac tgagtatgct
     5461 gttctgtcta tgtatgctct gtaagctacg agaaggactt ttttaaacag taaagggtgc
     5521 aatattttta caaattgaat taaataaagg ctgtctatta agtaatatgc ttgatatttt
     5581 tcttacttga tcggaaataa gaaaaaatat aaacgttgtt gctctaaaaa tcctagttca
     5641 gtttagccaa ccacaaatac ctttttgttc ctccaacagt ttttttttct tctctataat
     5701 atttggcagt ctatagtact ccaaatgttt ccccacagtc taactaattg gtacagccaa
     5761 aatcatgaca cttattgcaa taataatttt ggttcattgg cattgttgat agcctgtgcc
     5821 actaatatgg tcgattgatc atgcttcagg aagaaaacta tattgtttga tgtaagatta
     5881 ttaaatcttc acctgcttcc attacaaact attcccatct tattgaattc tggtatgtct
     5941 taaaggatta gttcacttcc caaatcaaaa tttacttagc cttttttcat ccatgatccc
     6001 tttttttcat cattaatgaa gaaattgttt ttgaaaaagt ttcaagattt ttttctctat
     6061 attgtggagc ttgttagttt aaaattccaa aatgcaatat gtggcttcaa atggttctaa
     6121 atgatcccag tcaaggaata acagtcttat ctaatgaaac cattagacct ttaaaaataa
     6181 aaaaaataaa agtatttatt tttaaatgac tgagtgatta agttgaattt cagcgtttcc
     6241 ttactgtgta gaagtccttc cttactggcc ccaccctttg gttctctgcc aatctgctac
     6301 ctaatgtaat gttgtggaac attattattc tttatttctt aattttttat tttttatttt
     6361 aaaacaatgt aaactgcaca gatgtgcagt ttgtttaaaa tggccaatgc tttggaaatg
     6421 catgacataa ttagatttca tgatgcacaa agccaaatct cagagcttgt gcaaaatgag
     6481 ctatcatttc actaggtaag accctaaatt ttcatatagg atcatttgga caattttgct
     6541 gcaggtaaaa tgcattctat agtccactgt cagccattgt tttggatagt atttattttt
     6601 ctctacaagt atagtcaata gttttctatt attttaaagg tttgtaacat ttaagggtga
     6661 ccaaatgcaa agtaaaattt cattttcggg tgaactatct cgtttaacat gggagaagtg
     6721 caaaacatac attattggct agaacattgt agtatttttt aaatggaaat gtgtgattgc
     6781 taatcttact ttgaatttgt ttacagggat cctttaaggg cgaattcgac ccaagtttgt
     6841 acaaaaaagc aggctggacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     6901 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     6961 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     7021 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     7081 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     7141 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     7201 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     7261 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     7321 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     7381 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     7441 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     7501 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     7561 tggacgagct gtacaagggc ggtggaagat ctgggaattc aaggcctctc gagcctctag
     7621 attctgcagc cctatagcac ccagctttct tgtacaaagt gggggatcca gacatgataa
     7681 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     7741 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     7801 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     7861 ccaactttat tatacatagt tgataattca ctggccgtcg ttttacggta cctctagaga
     7921 tccactagtg tcgacgatgt aggtcacggt ctcgaagccg cggtgcgggt gccagggcgt
     7981 gcccttg
//
