LOCUS Exported 7215 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FLAG-Stx17 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7215) AUTHORS Itakura E, Kishi-Itakura C, Mizushima N TITLE The hairpin-type tail-anchored SNARE syntaxin 17 targets to autophagosomes for fusion with endosomes/lysosomes. JOURNAL Cell. 2012 Dec 7;151(6):1256-69. doi: 10.1016/j.cell.2012.11.001. PUBMED 23217709 REFERENCE 2 (bases 1 to 7215) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7215 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..456 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(88..107) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 298..319 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 582..604 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 596..613 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 597..613 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 774..1153 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1154..1357 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1308..1328 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1387..1452 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 2430..3052 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2629..2648) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3076..3096) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3094..3391 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 3323..3342 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3445..4239 /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(3499..3518) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4109..4128 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4891..5025 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4928..4947) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4982..5001 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5030..5049) /label=T7 /note="T7 promoter, forward primer" promoter complement(5031..5049) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5063..5079) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5063..5079) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5076..5098) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5087..5103 /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(5111..5141) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5156..5177 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5294..5311) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5465..6053) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5545..5564) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6224..7084) /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 6847..6866 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7085..7189) /gene="bla" /label=AmpR promoter ORIGIN 1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 421 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa 481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg 541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 601 aacgacggcc agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt 661 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct 721 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg 781 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat 841 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 901 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 961 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 1021 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 1081 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat 1141 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga 1201 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 1261 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg 1321 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaatta 1381 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat 1441 gacgatgaca agcttgcggc cgcgaattca tcgatagatc cgatgtctga agatgaagaa 1501 aaagtgaaat tacgccgtct tgaaccagct atccagaaat tcattaagat agtaatccca 1561 acagacctgg aaaggttaag aaagcaccag ataaatattg agaagtatca aaggtgcaga 1621 atctgggaca agttgcatga agagcatatc aatgcaggac gtacagttca gcaactccga 1681 tccaatatcc gagaaattga gaaactttgt ttgaaagtcc gaaaggatga cctagtactt 1741 ctgaagagaa tgatagatcc tgttaaagaa gaagcatcag cagcaacagc agaatttctc 1801 caactccatt tggaatctgt agaagaactt aagaagcaat ttaatgatga agaaactttg 1861 ctacagcctc ctttgaccag atccatgact gttggtggag catttcatac tactgaagct 1921 gaagctagtt ctcagagttt gactcagata tatgccttac ctgaaattcc tcaagatcaa 1981 aatgctgcag aatcgtggga aaccttagaa gcggacttaa ttgaacttag ccaactggtc 2041 actgacttct ctctcctagt gaattctcag caggagaaga ttgacagcat tgcagaccat 2101 gtcaacagtg ctgctgtgaa tgttgaagag ggaaccaaaa acttagggaa ggctgcaaaa 2161 tacaagctgg cagctctgcc tgtggcaggt gcactcatcg ggggaatggt agggggtcct 2221 attggcctcc ttgcaggctt caaagtggca ggaattgcag ctgcacttgg tggtggggtg 2281 ttgggcttca caggtggaaa attgatacaa agaaagaaac agaaaatgat ggagaagctc 2341 acttccagct gtccagatct tcccagccaa actgacaaga aatgcagtta aggatctgat 2401 atcggtacca gtcgactcta gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc 2461 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa 2521 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg 2581 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac 2641 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag 2701 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca 2761 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca 2821 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt 2881 gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag caggaggacg 2941 tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc 3001 ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc 3061 ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3241 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3301 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 3361 tattccagaa gtagtgagga ggcttttttg gaggaattga tcagcttggg atctgatcaa 3421 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3481 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3541 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3601 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 3661 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 3721 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 3781 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 3841 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 3901 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 3961 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4021 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4081 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4141 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4201 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4261 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4321 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4381 gggatctcat gctggagttc ttcgcccacc ccgggctcga tcccctcgcg agttggttca 4441 gctgctgcct gaggctggac gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca 4501 tccaggaaac cagcagcggc tatccgcgca tccatgcccc cgaactgcag gagtggggag 4561 gcacgatggc cgctttggtc gacccggacg ggacgctcct gcgcctgata cagaacgaat 4621 tgcttgcagg catctcatga gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat 4681 ggagcgctgg cgcctgctgc gcgacggcga gctgctcacc acccactcgc caagctggaa 4741 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgccgatcat aatcagccat 4801 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 4861 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 4921 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4981 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaattc cctatagtga 5041 gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca 5101 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5161 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5221 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5281 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 6061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 6121 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 6181 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 6241 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 6301 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 6361 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6421 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6481 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6541 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6601 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6661 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6721 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6781 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6841 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6901 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6961 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 7021 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 7081 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 7141 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 7201 gaaaagtgcc acctg //