LOCUS       Exported                7488 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway destination vector with zebrafish aA-crystallin promoter 
            driving GFP in lens.
ACCESSION   .
VERSION     .
KEYWORDS    pDestTol2pACryGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7488)
  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 7488)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7488
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1056..1177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1300..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(1303..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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     2667..2683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2702..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(3514..4173)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4088..4107
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4227..4257)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(4282..4405)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4413..4429)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4445..4579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5251..5267)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5251..5267)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5304..5324)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5304..5322)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5343..5359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5356..5378)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5367..5383
                     /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(5391..5421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5436..5457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5574..5591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5745..6333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5825..5844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6504..7364)
                     /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     7127..7146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7365..7469)
                     /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 aggctcttgg 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 ccgccctgcc actcatcgca
     3541 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     3601 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     3661 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     3721 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     3781 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     3841 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     3901 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     3961 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     4021 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     4081 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     4141 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     4201 cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc
     4261 tggggtgcct aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta
     4321 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     4381 cgttcaactt tattatacat agttgataat tcactggccg tcgttttacg gtaccatcga
     4441 tgatgatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4501 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4561 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4621 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     4681 tgatcctcta gatcagatct gcgaagatac ggccacgggt gctcttgatc ctgtggctga
     4741 ttttggactg tgctgctcgc agctgctgat gaatcacata cttcctccat tttcttccac
     4801 tgattgactg ttataatttc cctaatttcc aggtcaaggt gctgtgcatt gtggtaatag
     4861 atgtgacatg acgtcacttc caaaggacca atgaacatgt ctgaccaatt tcatataatg
     4921 tgaaaacgat tttcataggc agaataaata acatttaaat taaactgggc atcagcgcaa
     4981 ttcaattggt ttggtaatag caagggaaaa tagaatgaag tgatctccaa aaaataagta
     5041 ctttttgact gtaaataaaa ttgtaaggag taaaaagtac ttttttttct aaaaaaatgt
     5101 aattaagtaa aagtaaaagt attgattttt aattgtactc aagtaaagta aaaatcccca
     5161 aaaataatac ttaagtacag taatcaagta aaattactca agtactttac acctctggtt
     5221 cttgaccccc taccttcagc aagcccagca gatccactag ttctagagcg gccgccaccg
     5281 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     5341 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5401 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5461 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5521 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6181 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtg
//
