LOCUS       Exported                7480 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway destination vector with zebrafish aA-crystallin promoter 
            driving mCherry GFP in lens.
ACCESSION   .
VERSION     .
KEYWORDS    pDESTtol2pACrymCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7480)
  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 7480)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7480
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1057..1178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1301..2011)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGHHSTGGMDELYK"
     primer_bind     complement(1381..1404)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1579..1598)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1848..1866
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2008..2017
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     2659..2675
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2694..2818
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2859..3164)
                     /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(2961..2980)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(3506..4165)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4080..4099
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4219..4249)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(4274..4397)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4405..4421)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4437..4571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5243..5259)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5243..5259)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5296..5316)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5296..5314)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5335..5351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5348..5370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5359..5375
                     /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(5383..5413)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5428..5449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5566..5583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5737..6325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5817..5836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6496..7356)
                     /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     7119..7138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7357..7461)
                     /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 aggcccattt acgccttaag atacattgat gagtttggac
     1081 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1141 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1201 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1261 aatgtggtat ggctgattat gatctagagt cgcggccgct ttacttgtac agctcgtcca
     1321 tgccgccggt ggagtggtgg ccctcggcgc gctcgtactg ttccacgatg gtgtagtcct
     1381 cgttgtggga ggtgatgtcc agcttgatgt tgacgttgta ggcgccgggc agctgcacgg
     1441 gcttcttggc cttgtaggtg gtcttgacct cggcgtcgta gtggccgccg tccttcagct
     1501 tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg
     1561 aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc
     1621 cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg
     1681 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
     1741 acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca
     1801 tgaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca
     1861 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
     1921 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg
     1981 ccatgttgtc atcctcgccc ttgctcacca tggtggcgac cggtaatgtc agacctggta
     2041 actccttact gtaacttatt gcacctgatg aacaaaagat gtgaactgaa cgcaagtagg
     2101 aggctaatag aggctagtac agtacaaaga tccccaaatg atgtgtgccg gccttagtgg
     2161 aggcagaccc ctggttatat acctgtgctg cagaaagagg ggcttacaaa tggatctctc
     2221 tggaatggca ttatctcatg gtgggaacag tgttgtcagc agaatcagcg gggtggactc
     2281 tgacctgaga atgacagtct agcactgaat ggctcagaca atactcttgt ggccaactat
     2341 caccggaaaa taagtgcctg ccaaaacaaa ccagaccaga tgaagctaca tggctgtaga
     2401 acgtcagtat ttaatgtgtt gtggccactt ggttcaccag atttaagagc taattttttt
     2461 aaatatcaca gataatcaat cgcttttttt tttcttgact gaaaagctta gactggaaaa
     2521 ttctttgaaa aagtttgcac tacagaaaat aaatacataa catcctgcac tgaatgcaca
     2581 ctattaatag aaaagagagc attattgtag gaaatgtatt gtatatgtac attctgtgca
     2641 tttcacaatt ttctcgagca ggaaacagct atgaccatga ttacgccaag ctatcaactt
     2701 tgtatagaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     2761 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggt
     2821 cgacctgcag actggctgtg tataagggag cctgacattt atattcccca gaacatcagg
     2881 ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata
     2941 acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata
     3001 tgcaccaccg ggtaaagttc acgggggact ttatctgaca gcagacgtgc actggccagg
     3061 gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc cacaaacaga
     3121 cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc agcccctgtt
     3181 ctcgtcggca aaggagccgt tcatttcaat aaaccgggcg acctcagcca tcccttcctg
     3241 attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca tggttgtgct
     3301 taccgaaccg gagatattga catcatatat gccttgagca actgatagct gtcgctgtca
     3361 actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt cgggtataca
     3421 tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg
     3481 cttttagtaa gccggatcct ctagattacg ccccgccctg ccactcatcg cagtactgtt
     3541 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     3601 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     3661 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     3721 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     3781 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     3841 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     3901 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
     3961 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     4021 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     4081 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     4141 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacggat
     4201 cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc
     4261 ctaatgcggc cgccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt
     4321 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac
     4381 tttattatac atagttgata attcactggc cgtcgtttta cggtaccatc gatgatgatc
     4441 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     4501 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4561 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     4621 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcctc
     4681 tagatcagat ctgcgaagat acggccacgg gtgctcttga tcctgtggct gattttggac
     4741 tgtgctgctc gcagctgctg atgaatcaca tacttcctcc attttcttcc actgattgac
     4801 tgttataatt tccctaattt ccaggtcaag gtgctgtgca ttgtggtaat agatgtgaca
     4861 tgacgtcact tccaaaggac caatgaacat gtctgaccaa tttcatataa tgtgaaaacg
     4921 attttcatag gcagaataaa taacatttaa attaaactgg gcatcagcgc aattcaattg
     4981 gtttggtaat agcaagggaa aatagaatga agtgatctcc aaaaaataag tactttttga
     5041 ctgtaaataa aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt
     5101 aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat
     5161 acttaagtac agtaatcaag taaaattact caagtacttt acacctctgg ttcttgaccc
     5221 cctaccttca gcaagcccag cagatccact agttctagag cggccgccac cgcggtggag
     5281 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     5341 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5401 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5461 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5521 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5581 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5641 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5701 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5761 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5821 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5881 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5941 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6001 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6061 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6121 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6181 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6781 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7081 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
//
