LOCUS       sequence-392889-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector with zebrafish aA-crystallin promoter 
            driving GFP in lens.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Berger J, Currie PD
  TITLE     503unc, a small and muscle-specific zebrafish promoter.
  JOURNAL   Genesis. 2013 Jun;51(6):443-7. doi: 10.1002/dvg.22385. Epub 2013 Mar
            26.
  PUBMED    23444339
REFERENCE   2  (bases 1 to 7487)
  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/dvg.22385";
            journalName: "Genesis"; date: "2013-06"; volume: "51"; issue: "6"; 
            pages: "443-7"
COMMENT     Sequence Label: pDestTol2pACryGFP
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    1056..1177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(1300..2013,2014..2016,2017..2019))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1303..2013,2014..2016,2017..2019))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1345..1366)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1694..1713
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1953..1974
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2016..2025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2667..2683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2703..2826
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2867..3172)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2969..2988)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(3492..4172)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     4087..4106
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(4226..4232,4233..4250,4251..4256))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(4281..4404)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4412..4428)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4444..4578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(5017..5204,5205..5216)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5250..5266)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5250..5266)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5303..5323)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5303..5321)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5342..5358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5355..5377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5366..5382
                     /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(5390..5396,5397..5414,5415..5420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5435..5456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5573..5590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5744..6332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5824..5843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6503..7294,7295..7363))
                     /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     7126..7145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7364..7468)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatt ggccttaaga tacattgatg agtttggaca
     1081 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1141 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1201 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1261 atgtggtatg gctgattatg atctagagtc gcggccgctt tacttgtaca gctcgtccat
     1321 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     1381 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
     1441 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     1501 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     1561 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     1621 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1681 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     1741 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1801 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     1861 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     1921 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1981 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtaatgtcag
     2041 acctggtaac tccttactgt aacttattgc acctgatgaa caaaagatgt gaactgaacg
     2101 caagtaggag gctaatagag gctagtacag tacaaagatc cccaaatgat gtgtgccggc
     2161 cttagtggag gcagacccct ggttatatac ctgtgctgca gaaagagggg cttacaaatg
     2221 gatctctctg gaatggcatt atctcatggt gggaacagtg ttgtcagcag aatcagcggg
     2281 gtggactctg acctgagaat gacagtctag cactgaatgg ctcagacaat actcttgtgg
     2341 ccaactatca ccggaaaata agtgcctgcc aaaacaaacc agaccagatg aagctacatg
     2401 gctgtagaac gtcagtattt aatgtgttgt ggccacttgg ttcaccagat ttaagagcta
     2461 atttttttaa atatcacaga taatcaatcg cttttttttt tcttgactga aaagcttaga
     2521 ctggaaaatt ctttgaaaaa gtttgcacta cagaaaataa atacataaca tcctgcactg
     2581 aatgcacact attaatagaa aagagagcat tattgtagga aatgtattgt atatgtacat
     2641 tctgtgcatt tcacaatttt ctcgagcagg aaacagctat gaccatgatt acgccaagct
     2701 atcaactttg tatagaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
     2761 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     2821 actatggtcg acctgcagac tggctgtgta taagggagcc tgacatttat attccccaga
     2881 acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt
     2941 ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat
     3001 ccccgatatg caccaccggg taaagttcac gggggacttt atctgacagc agacgtgcac
     3061 tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca
     3121 caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag
     3181 cccctgttct cgtcggcaaa ggagccgttc atttcaataa accgggcgac ctcagccatc
     3241 ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc attctgcatg
     3301 gttgtgctta ccgaaccgga gatattgaca tcatatatgc cttgagcaac tgatagctgt
     3361 cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg acatacttcg
     3421 ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg
     3481 ttatctggct tttagtaagc cggatcctct agattacgcc ccgcctgcca ctcatcgcag
     3541 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     3601 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     3661 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     3721 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     3781 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     3841 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     3901 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc
     3961 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     4021 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     4081 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     4141 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     4201 gacggatcct aactcaaaat ccacacatta tacgagccgg aagcataaag tgtaaagcct
     4261 ggggtgccta atgcggccgc catagtgact ggatatgttg tgttttacag tattatgtag
     4321 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     4381 gttcaacttt attatacata gttgataatt cactggccgt cgttttacgg taccatcgat
     4441 gatgatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4501 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4561 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4621 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     4681 gatcctctag atcagatctg cgaagatacg gccacgggtg ctcttgatcc tgtggctgat
     4741 tttggactgt gctgctcgca gctgctgatg aatcacatac ttcctccatt ttcttccact
     4801 gattgactgt tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga
     4861 tgtgacatga cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt catataatgt
     4921 gaaaacgatt ttcataggca gaataaataa catttaaatt aaactgggca tcagcgcaat
     4981 tcaattggtt tggtaatagc aagggaaaat agaatgaagt gatctccaaa aaataagtac
     5041 tttttgactg taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta
     5101 attaagtaaa agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa
     5161 aaataatact taagtacagt aatcaagtaa aattactcaa gtactttaca cctctggttc
     5221 ttgaccccct accttcagca agcccagcag atccactagt tctagagcgg ccgccaccgc
     5281 ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     5341 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5521 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6181 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6361 ctgacgctca gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga ttatcaaaaa
     6421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6781 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6841 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6901 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6961 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7021 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7081 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7141 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7201 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7261 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7321 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7381 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7441 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtg
//
