LOCUS Exported 5635 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of zebrafish GFP-tagged Rab26 in mammalian cells.. ACCESSION . VERSION . KEYWORDS EGFP-Rab26 in pCSDEST2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5635) TITLE New_for_revision JOURNAL Unpublished REFERENCE 2 (bases 1 to 5635) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5635 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 168..1145 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1180..1198 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1180..1197 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 1253..1277 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 1278..1287 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1284..2000 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1284..2000 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1329..1350) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1590..1609) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1937..1958 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(2002..2026) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2772..2792) /gene="mutant version of attB" /label=attB3 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" promoter complement(2810..2827) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2832..2966 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2856..2875) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2910..2929 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3079..3099) /label=T3 /note="T3 promoter, forward primer" promoter complement(3079..3097) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3118..3134) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3118..3134) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3131..3153) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3142..3158 /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(3166..3196) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3211..3232 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3349..3366) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3520..4108) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3600..3619) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4279..5139) /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 4902..4921 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5140..5244) /gene="bla" /label=AmpR promoter rep_origin complement(5270..90) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5407..5428) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5619..3 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 61 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt 121 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat 181 ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat 241 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat 301 ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag 361 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc 421 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata 481 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat 541 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc 601 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac 661 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg 721 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg 781 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat 841 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact 901 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg 961 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct 1021 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat 1081 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag 1141 ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat 1201 acaagctact tgttcttttt gcaggatccc atcgattcga attcaaggat caacaagttt 1261 gtacaaaaaa gcaggctgcc accatggtga gcaagggcga ggagctgttc accggggtgg 1321 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 1381 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 1441 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 1501 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 1561 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 1621 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1681 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1741 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 1801 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 1861 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 1921 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 1981 gcatggacga gctgtacaag cacccagctt tcttgtacaa agtggtgtcg aggaaaaaag 2041 cgctgaaggg gaagggcagt tcaaacacac cgaattcatc tccgagcgct gaaaaagaga 2101 aggggaaaaa agtgtcgagc caggcgaacg gagtggttca tcccagccgg ccatctctca 2161 gcaacagcgg ggagttctac gacatcgcat tcaaggtgat gctggtcgga gactctggag 2221 tggggaaaac ctgtctgctg gttcgcttta aagatggagc atttctggcc ggaagcttca 2281 tctcaactgt tggcatagac ttcaggaata aagtgctgag catcgacagc atgagggtta 2341 aactacagat ctgggataca gcgggacagg agcgtttcag gagtgtgacg cacgcatact 2401 atcgagacgc tcacgctctt ttgctgcttt atgatgtgac aaataaatcc tcctttgaca 2461 acatacaggc ctggctgacg gagattcacg agttcgctca gcagaacgtg gtcattatgc 2521 tgttgggtaa taaggctgat gctacacatg aacgcattgt gaagagggag gagggggaga 2581 aactagctaa ggagtttggc gttcccttca tggagaccag tgctaaaaca ggcctgaatg 2641 tggagttggc cttcactgcg gttgccaagg aactgaagca cagggagtta aaggaaccaa 2701 acgagcccaa gtttcagctg caggagtacg tgaacaagga gatgaaggga gtcgggtgct 2761 gtcgctcata acaactttat tatacatagt tgcactcgag cctctagaac tatagtgagt 2821 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta 2881 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 2941 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 3001 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc 3061 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc 3121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 3181 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 3241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 3361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 3421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 3481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 3541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 3601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 3661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 3721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 3781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 3841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 3901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 3961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 4081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 4261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 4321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 4381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 4441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 4501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 4561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 4621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 4681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 4741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 4801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 4861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 4921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 4981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 5041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 5101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 5161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 5221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc 5281 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 5341 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 5401 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 5461 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 5521 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 5581 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaag //