LOCUS Exported 7479 bp ds-DNA circular SYN 23-SEP-2022 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 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 Sep 23, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7479 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(18..34) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" regulatory 676..685 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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" primer_bind 2528..2549 /label=F1ori-F /note="F1 origin, forward primer" promoter 2712..2816 /gene="bla" /label=AmpR promoter CDS 2817..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 4760..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" 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 5924..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..7478) /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 //