LOCUS Exported 5575 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of zebrafish GFP-tagged Rab33bb in mammalian cells.. ACCESSION . VERSION . KEYWORDS EGFP-Rab33bb in pCSDEST2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5575) TITLE New_for_revision JOURNAL Unpublished REFERENCE 2 (bases 1 to 5575) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5575 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 108..132 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 133..142 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 139..855 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 139..855 /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(184..205) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(445..464) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 792..813 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(857..881) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1564..1584) /gene="mutant version of attB" /label=attB3 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" promoter complement(1602..1619) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1624..1758 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1648..1667) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1702..1721 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1871..1891) /label=T3 /note="T3 promoter, forward primer" promoter complement(1871..1889) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1910..1926) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1910..1926) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1923..1945) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1934..1950 /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(1958..1988) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2003..2024 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2141..2158) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2312..2900) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2392..2411) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3071..3931) /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 3694..3713 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3932..4036) /gene="bla" /label=AmpR promoter rep_origin complement(4062..4517) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4199..4220) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4411..4430 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4591..5575 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca 121 aaaaagcagg ctgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 181 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 241 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 301 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc 361 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 421 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 481 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 541 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg 601 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 661 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 721 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag 781 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 841 gacgagctgt acaagcaccc agctttcttg tacaaagtgg tggattcgtc tttagaaagc 901 tccagctcct ctaacggccc cccgacgcgc tgctgtcgga cctgtaagat catcgtgatc 961 ggggatgccg gtgtgggcaa gacctgtctg acacatcgat tctgcaccgg acaatttcct 1021 accagaacag aggcgaccat cggggtggat tttcgagaga aacttcttca gatagacggc 1081 gaaaaaataa aagtacagct atgggacacg gccggtcagg agcgcttccg taaaagcatg 1141 gtgcagcact actatcgaaa tgtgcatggt atcgtcttcg tctttgacgt cacaaaccaa 1201 tccagtttct gcaatctgcc tttgtggatg gacgagtgcc gtcagcactc tcttggtctg 1261 gaaattccac gtgccctagt ttgcaacaag gctgacctga tgtcttcaaa tgcttctgtg 1321 ctaatcgagc aggctcgtca tttggctgaa gtccacggga tgtcttttta tctcacttca 1381 gccaaaggtc ttaaaggcga ccaggtggac agcatcttca tggctttagc acagaggctg 1441 aagagccaga gaagggagag cgtaaatgga cgtgtgctag agtgcagagc tgagagctta 1501 aaaatccagg cacgggttat aacagcccaa gagactaaaa agaaatggac ctgcatctgc 1561 taacaacttt attatacata gttgcactcg agcctctaga actatagtga gtcgtattac 1621 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag 1681 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata 1741 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg 1801 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca 1861 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt 1921 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 1981 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 2041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 2101 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 2161 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 2221 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 2281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 2341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 2401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 2461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 2521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 2581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 2641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 2701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 2761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 2821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 2881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 2941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 3001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 3061 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 3121 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 3181 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 3241 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 3301 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 3361 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 3421 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 3481 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 3541 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 3601 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 3661 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 3721 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 3781 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 3841 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 3901 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 3961 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 4021 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat 4081 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 4141 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 4201 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 4261 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 4321 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 4381 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 4441 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 4501 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc 4561 ggtgcgggcc tcttcgctat tacgccagtc gaccatagcc aattcaatat ggcgtatatg 4621 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg 4681 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg 4741 gcaccatgcc aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt 4801 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt 4861 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat 4921 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc 4981 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg 5041 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg 5101 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac 5161 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc 5221 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca 5281 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca 5341 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca 5401 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca 5461 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat 5521 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac //