LOCUS       sequence-372953-        7479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector with zebrafish aA-crystallin promoter 
            driving mCherry GFP in lens.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  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 7479)
  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: pDESTtol2pACrymCherry
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..34)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      676..685
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..1392
                     /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(827..845)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1095..1114
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1289..1312
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1515..1636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2005..2167,2168..2179)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     2528..2549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2712..2816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2817..2885,2886..3677)
                     /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     complement(3035..3054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3848..4436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4337..4356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4590..4607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4724..4745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4760..4765,4766..4783,4784..4790)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4798..4814
                     /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)."
     primer_bind     4803..4825
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4822..4838
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4857..4877
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4859..4877
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     4914..4930
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4914..4930
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(4964..4975,4976..5163)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5602..5736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5752..5768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5776..5899
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5924..5929,5930..5947,5948..5954)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6008..6688
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(6074..6093)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             7008..7313
                     /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     7192..7211
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7354..7477)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 agcttggcgt aatcatggtc atagctgttt cctgctcgag aaaattgtga aatgcacaga
       61 atgtacatat acaatacatt tcctacaata atgctctctt ttctattaat agtgtgcatt
      121 cagtgcagga tgttatgtat ttattttctg tagtgcaaac tttttcaaag aattttccag
      181 tctaagcttt tcagtcaaga aaaaaaaaag cgattgatta tctgtgatat ttaaaaaaat
      241 tagctcttaa atctggtgaa ccaagtggcc acaacacatt aaatactgac gttctacagc
      301 catgtagctt catctggtct ggtttgtttt ggcaggcact tattttccgg tgatagttgg
      361 ccacaagagt attgtctgag ccattcagtg ctagactgtc attctcaggt cagagtccac
      421 cccgctgatt ctgctgacaa cactgttccc accatgagat aatgccattc cagagagatc
      481 catttgtaag cccctctttc tgcagcacag gtatataacc aggggtctgc ctccactaag
      541 gccggcacac atcatttggg gatctttgta ctgtactagc ctctattagc ctcctacttg
      601 cgttcagttc acatcttttg ttcatcaggt gcaataagtt acagtaagga gttaccaggt
      661 ctgacattac cggtcgccac catggtgagc aagggcgagg atgacaacat ggccatcatc
      721 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      781 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      841 accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
      901 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
      961 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1021 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1081 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1141 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     1201 gacggcggcc actacgacgc cgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1261 ctgcccggcg cctacaacgt caacatcaag ctggacatca cctcccacaa cgaggactac
     1321 accatcgtgg aacagtacga gcgcgccgag ggccaccact ccaccggcgg catggacgag
     1381 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     1441 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     1501 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1561 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1621 tcatcaatgt atcttaaggc gtaaatgggc ctgtgtttca gacaccaggg agtctctgct
     1681 cacgtttcct gctatttgca gcctctctat caagactaat acacctcttc ccgcatcggc
     1741 tgcctgtgag aggcttttca gcactgcagg attgcttttc agccccaaaa gagctaggct
     1801 tgacactaac aattttgaga atcagcttct actgaagtta aatctgaggt tttacaactt
     1861 tgagtagcgt gtactggcat tagattgtct gtcttatagt ttgataatta aatacaaaca
     1921 gttctaaagc aggataaaac cttgtatgca tttcatttaa tgttttttga gattaaaagc
     1981 ttaaacaaga atctctagtt ttctttcttg cttttacttt tacttcctta atactcaagt
     2041 acaattttaa tggagtactt ttttactttt actcaagtaa gattctagcc agatactttt
     2101 acttttaatt gagtaaaatt ttccctaagt acttgtactt tcacttgagt aaaatttttg
     2161 agtacttttt acacctctgt caagaactcc tggacaaacc tctgacctgt gtggaacaga
     2221 gtggatatgg gtgtctgaac agatattcac gtcttttgca gatcagaggg catttctggt
     2281 gttgtgcaat gacctgggtc caacaggtag gcgtactcgt atccaagagc attgttggac
     2341 aggaggtagt ctccattcct ataaagagga aggaatggca ccatgtagta gccgtcattg
     2401 tggccaattg gtgcattggt acgtgggtag atggaccggg gaggctgatg agttcttagc
     2461 cagcgtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2581 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2641 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2701 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2761 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3001 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3361 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3781 aaatgcctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3961 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4561 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4621 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4681 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     4741 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     4801 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     4861 ttaaccctca ctaaagggaa caaaagctgg agctccaccg cggtggcggc cgctctagaa
     4921 ctagtggatc tgctgggctt gctgaaggta gggggtcaag aaccagaggt gtaaagtact
     4981 tgagtaattt tacttgatta ctgtacttaa gtattatttt tggggatttt tactttactt
     5041 gagtacaatt aaaaatcaat acttttactt ttacttaatt acattttttt agaaaaaaaa
     5101 gtacttttta ctccttacaa ttttatttac agtcaaaaag tacttatttt ttggagatca
     5161 cttcattcta ttttcccttg ctattaccaa accaattgaa ttgcgctgat gcccagttta
     5221 atttaaatgt tatttattct gcctatgaaa atcgttttca cattatatga aattggtcag
     5281 acatgttcat tggtcctttg gaagtgacgt catgtcacat ctattaccac aatgcacagc
     5341 accttgacct ggaaattagg gaaattataa cagtcaatca gtggaagaaa atggaggaag
     5401 tatgtgattc atcagcagct gcgagcagca cagtccaaaa tcagccacag gatcaagagc
     5461 acccgtggcc gtatcttcgc agatctgatc tagaggatca taatcagcca taccacattt
     5521 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     5581 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5641 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5701 tccaaactca tcaatgtatc ttatcatgtc tggatcatca tcgatggtac cgtaaaacga
     5761 cggccagtga attatcaact atgtataata aagttgaacg agaaacgtaa aatgatataa
     5821 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     5881 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     5941 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
     6001 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     6061 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     6121 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     6181 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     6241 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     6301 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     6361 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     6421 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     6481 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     6541 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     6601 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcaggcggg
     6661 gcgtaatcta gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
     6721 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
     6781 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     6841 catatatgat gtcaatatct ccggttcggt aagcacaacc atgcagaatg aagcccgtcg
     6901 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     6961 tattgaaatg aacggctcct ttgccgacga gaacaggggc tggtgaaatg cagtttaagg
     7021 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     7081 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     7141 aagtcccccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     7201 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
     7261 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
     7321 gctcccttat acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta
     7381 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     7441 ttttacgttt ctcgttcaac ttttctatac aaagttgat
//
