LOCUS       sequence-227691-        8786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8786)
  AUTHORS   Mosimann C, Kaufman CK, Li P, Pugach EK, Tamplin OJ, Zon LI
  TITLE     Ubiquitous transgene expression and Cre-based recombination driven 
            by the ubiquitin promoter in zebrafish.
  JOURNAL   Development. 2011 Jan . 138(1):169-77.
  PUBMED    21138979
REFERENCE   2  (bases 1 to 8786)
  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: 
            "Development. 2011 Jan . 138(1):169-77."
COMMENT     Sequence Label: pDestTol2pA2_ubi:EGFP
FEATURES             Location/Qualifiers
     source          1..8786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..37)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    53..187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(626..813,814..825)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(859..875)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(859..875)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(912..932)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(912..930)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(951..967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(964..986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    975..991
                     /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(999..1005,1006..1023,1024..1029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1044..1065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1182..1199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1353..1941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1433..1452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2112..2903,2904..2972))
                     /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     2735..2754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2973..3077)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3240..3261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(3610..3621,3622..3784)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     4157..4173
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4193..4213
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    7748..7772
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(7778..7780,7781..7783,7784..8494)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7778..7780,7781..7783,7784..8494)
                     /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(7823..7844)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8084..8103)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8431..8452
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(8556..8580)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    8582..8716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(join(8779..8786,1..13))
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 ttatacatag ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga
       61 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      121 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      181 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      241 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
      301 tcagatctgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg
      361 ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt
      421 ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac
      481 gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt
      541 tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt
      601 ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt
      661 aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa
      721 gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt
      781 aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta
      841 ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
      901 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
      961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1981 tggaacgaaa actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc
     2041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     3121 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3181 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3241 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3301 ccgtctatca gggcgatggc ccacgctggc taagaactca tcagcctccc cggtccatct
     3361 acccacgtac caatgcacca attggccaca atgacggcta ctacatggtg ccattccttc
     3421 ctctttatag gaatggagac tacctcctgt ccaacaatgc tcttggatac gagtacgcct
     3481 acctgttgga cccaggtcat tgcacaacac cagaaatgcc ctctgatctg caaaagacgt
     3541 gaatatctgt tcagacaccc atatccactc tgttccacac aggtcagagg tttgtccagg
     3601 agttcttgac agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac
     3661 ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa
     3721 aagtaaaaaa gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca
     3781 agaaagaaaa ctagagattc ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg
     3841 catacaaggt tttatcctgc tttagaactg tttgtattta attatcaaac tataagacag
     3901 acaatctaat gccagtacac gctactcaaa gttgtaaaac ctcagattta acttcagtag
     3961 aagctgattc tcaaaattgt tagtgtcaag cctagctctt ttggggctga aaagcaatcc
     4021 tgcagtgctg aaaagcctct cacaggcagc cgatgcggga agaggtgtat tagtcttgat
     4081 agagaggctg caaatagcag gaaacgtgag cagagactcc ctggtgtctg aaacacaggc
     4141 cagatgggcc ctcgagcagg aaacagctat gaccatgatt acgccaagct atcaactttg
     4201 tatagaaaag ttggctccga attcgccctt aaaactcgag accagcaaag ttctagaatt
     4261 tgtcgaaaca tttatgttat atatttcctg aaaaaaattc tgagtaagtt cttaagtgtt
     4321 attgccagca acataaacaa cagacggcaa aatgaataaa tgataacaaa gcagtaggct
     4381 taaataaacc taatttttat aggctgttct ctacaaccct caaacagtga ttagttttgt
     4441 acttataaac ttgccctttc attcatattt caagaaaatt ggttcagaag atctggatat
     4501 tctagcagtt gttcaagctc atggagggat cagtgacctg attccacaat gactaggcct
     4561 aatccagaaa ttagatgact gtcaacataa aaaggcacag cactcactag ctgccctata
     4621 tattttatta tattttacat atattatttt atttatttag ctctgagtgc tgtactttct
     4681 ggttaaagaa aactgcttac aacagctaac ctgtactacc tcaggctcag ggaatttgga
     4741 acaggtttgt ctggtttgtt tctttaacca tgcatgcttg ttttcaacta tggcaacaca
     4801 gtcacatggg acattacaga aatgatttgt cgatgacatg cgacttttct ttaataaagc
     4861 gcaaagatcc caaaaagcaa acttttaaca aaaatcatat aattatattt tcaatccagc
     4921 tttgtagcaa ctttgtgctg ctgttcactc agcaacagat agtcagtata aggtcagtgt
     4981 gtctcaaagc agtgccatct gtttcacaca ttgcgttcta tatataagtg tgctggttga
     5041 cacgacactg tataaggcct aggctaaaac acaaacaatg tagaatgaca ctgtgttttt
     5101 tttgtaaaca aatgttgttt ttggttaaac atctttgtga aaacatcctc ctgtcatgta
     5161 tttgctatat tcaaatgtta aacccgtgca gaatagaaca tatacaaaaa aaaacaacac
     5221 aacacatttt taaacattat taaatatcaa gtattgctgg cagttctgtt tctgttttac
     5281 agtacccttt gccacagttc tccgcttttc ctggtccaga ttccacaagt ctgattcacc
     5341 aatagcaaag cgaataaaca accaaagcag ccaatcactg cttgtagact gtcctgcgag
     5401 accggcccat tccagcacat tctggaaact tcctttatat gataattata aatacattta
     5461 aattattgat acaaaacatg taattcctag aacataacca tagcaatcat tagttttcag
     5521 ggtaattatg tatttttagg atttgactgc ggaaagatct ggtcatgtga cgtctcatga
     5581 acgtcacggc cctgggtttc tataaataca gtaggactct cgaccatcgg cagatttttc
     5641 gaagaagaag atcagtttca ggagccgtac tgttccgttt caacgcaaat attaacggta
     5701 agagcgaatt tcctagtttg ttttcatgcc attctttaaa accatagcgt attactttaa
     5761 ttatagtaaa ctttcgcttt ctttattaca agagacgttt tgtgttgatt ctccgcggac
     5821 attttcggtc agacaatcag aaaatgaccg cggaggacca gtaacttgca ttacacgtaa
     5881 gttaaatctt cgtgtattaa aatggttagg ttgttaacgt caaataggtt accgtgtttg
     5941 cgtgtgatca ggttggtttt gttagatttt tgtcagtatt tttaatttat ttgttttagt
     6001 ttatttattt tttttgctga atcatagttt gtgaacaaag aacccggatg ttacatacag
     6061 tacagccgcc atgttacaga gagttataac ttaatcattt taaaaataat tttgccttac
     6121 ttttagtttg tcatgttgag aaatgaggaa atgttaaaat gaggaaatat ccaattaatt
     6181 taatatatca aaataatcca tgattacaat gcactgaact ggagaaaatt aagatgtttt
     6241 ctagtgtcat gaaacaaatg taagagatgt acattgtaga tgttttatgt caagaattgg
     6301 ctagttgatg cagcatactg gcgatactca gttgtaataa cagtaacgtt acatgttaat
     6361 agactactga gtatgctgtt ctgtctatgt atgctctgta agctacgaga aggacttttt
     6421 taaacagtaa agggtgcaat atttttacaa attgaattaa ataaaggctg tctattaagt
     6481 aatatgcttg atatttttct tacttgatcg gaaataagaa aaaatataaa cgttgttgct
     6541 ctaaaaatcc tagttcagtt tagccaacca caaatacctt tttgttcctc caacagtttt
     6601 tttttcttct ctataatatt tggcagtcta tagtactcca aatgtttccc cacagtctaa
     6661 ctaattggta cagccaaaat catgacactt attgcaataa taattttggt tcattggcat
     6721 tgttgatagc ctgtgccact aatatggtcg attgatcatg cttcaggaag aaaactatat
     6781 tgtttgatgt aagattatta aatcttcacc tgcttccatt acaaactatt cccatcttat
     6841 tgaattctgg tatgtcttaa aggattagtt cacttcccaa atcaaaattt acttagcctt
     6901 ttttcatcca tgatcccttt ttttcatcat taatgaagaa attgtttttg aaaaagtttc
     6961 aagatttttt tctctatatt gtggagcttg ttagtttaaa attccaaaat gcaatatgtg
     7021 gcttcaaatg gttctaaatg atcccagtca aggaataaca gtcttatcta atgaaaccat
     7081 tagaccttta aaaataaaaa aaataaaagt atttattttt aaatgactga gtgattaagt
     7141 tgaatttcag cgtttcctta ctgtgtagaa gtccttcctt actggcccca ccctttggtt
     7201 ctctgccaat ctgctaccta atgtaatgtt gtggaacatt attattcttt atttcttaat
     7261 tttttatttt ttattttaaa acaatgtaaa ctgcacagat gtgcagtttg tttaaaatgg
     7321 ccaatgcttt ggaaatgcat gacataatta gatttcatga tgcacaaagc caaatctcag
     7381 agcttgtgca aaatgagcta tcatttcact aggtaagacc ctaaattttc atataggatc
     7441 atttggacaa ttttgctgca ggtaaaatgc attctatagt ccactgtcag ccattgtttt
     7501 ggatagtatt tatttttctc tacaagtata gtcaatagtt ttctattatt ttaaaggttt
     7561 gtaacattta agggtgacca aatgcaaagt aaaatttcat tttcgggtga actatctcgt
     7621 ttaacatggg agaagtgcaa aacatacatt attggctaga acattgtagt attttttaaa
     7681 tggaaatgtg tgattgctaa tcttactttg aatttgttta cagggatcct ttaagggcga
     7741 attcgaccca agtttgtaca aaaaagcagg ctggaccatg gtgagcaagg gcgaggagct
     7801 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     7861 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     7921 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     7981 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     8041 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     8101 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     8161 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     8221 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     8281 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     8341 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     8401 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     8461 cgggatcact ctcggcatgg acgagctgta caagggcggt ggaagatctg ggaattcaag
     8521 gcctctcgag cctctagatt ctgcagccct atagcaccca gctttcttgt acaaagtggg
     8581 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     8641 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     8701 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     8761 ggtgtgggag gttttttcca acttta
//
