LOCUS Exported 6780 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS EGFP-Rab7A T22N SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6780) AUTHORS Sun Q, Westphal W, Wong KN, Tan I, Zhong Q TITLE Rubicon controls endosome maturation as a Rab7 effector. JOURNAL Proc Natl Acad Sci U S A. 2010 Nov 9. 107(45):19338-43. PUBMED 20974968 REFERENCE 2 (bases 1 to 6780) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6780 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(42..61) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 233..612 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 613..816 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 767..787 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 861..880 /label=T7 /note="T7 promoter, forward primer" promoter 861..879 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 893..1609 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 893..1609 /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(938..959) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1199..1218) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1546..1567 /label=EGFP-C /note="EGFP, forward primer" CDS 1619..1645 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1646..1672 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(2331..2349) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2332..2349) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2369..2386) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2375..2599 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2645..3073 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2732..2751) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2942..2963 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3082..3102) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3087..3416 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3267..3402 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3329..3348 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3483..4277 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3537..3556) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4147..4166 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4451..4572 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4488..4507) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4542..4561 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4621..4637) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4621..4637) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4634..4656) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4645..4661 /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(4669..4699) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4714..4735 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4852..4869) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5023..5611) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5103..5122) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5782..6642) /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 6405..6424 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6643..6747) /gene="bla" /label=AmpR promoter ORIGIN 1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact 841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttatggtgag 901 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 961 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 1021 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 1081 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 1141 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 1201 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 1261 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 1321 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 1381 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 1441 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 1501 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 1561 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gatccatgta 1621 cccatacgat gttccggatt acgcttaccc atacgatgtt ccggattacg ctggtacctc 1681 taggaagaaa gtgttgctga aggttatcat cctgggagat tctggagtcg ggaagaactc 1741 actcatgaac cagtatgtga ataagaaatt cagcaatcag tacaaagcca caataggagc 1801 tgactttctg accaaggagg tgatggtgga tgacaggcta gtcacaatgc agatatggga 1861 cacagcagga caggaacggt tccagtctct cggtgtggcc ttctacagag gtgcagactg 1921 ctgcgttctg gtatttgatg tgactgcccc caacacattc aaaaccctag atagctggag 1981 agatgagttt ctcatccagg ccagtccccg agatcctgaa aacttcccat ttgttgtgtt 2041 gggaaacaag attgacctcg aaaacagaca agtggccaca aagcgggcac aggcctggtg 2101 ctacagcaaa aacaacattc cctactttga gaccagtgcc aaggaggcca tcaacgtgga 2161 gcaggcgttc cagacgattg cacggaatgc acttaagcag gaaacggagg tggagctgta 2221 caacgaattt cctgaaccta tcaaactgga caagaatgac cgggccaagg cctcggcaga 2281 aagctgcagt tgctaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt 2341 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 2401 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 2461 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 2521 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 2581 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 2641 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2701 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2761 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 2821 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 2881 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 2941 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3001 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 3061 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3421 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 3481 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 3541 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 3601 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3661 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3721 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 3781 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 3841 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 3901 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3961 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4021 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 4081 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4141 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 4201 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 4261 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 4321 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 4381 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 4441 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 4501 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 4561 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 4621 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4681 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 4801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 4921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 5041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 5221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 5281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 5341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 5581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 5641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 5701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 5761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 5821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 5881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 5941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 6001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 6061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 6121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 6181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 6241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 6301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 6361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 6421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 6481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 6541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 6601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 6661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 6721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga //