LOCUS V030576 7407 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030576 VERSION V030576 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1029..3545 /label="Atg9a(NM_001003917)" /note="Atg9a(NM_001003917)" /gene="Atg9a" CDS 3573..3602 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 3621..3644 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(3664..3680) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 3691..4313 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(4462..5050) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 5379..5426 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(5658..6452) /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" promoter complement(6487..6844) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(6846..6950) /gene="bla" /label="AmpR promoter" rep_origin 6977..7405 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat ggcacagttt gacactgaat accagcgcct agaggcctcc tatagcgatt 1081 cgccccccgg ggaagaggac ctgttggtgc atgtggccga ggggagcaaa tcaccttggc 1141 accacattga aaaccttgac ctcttcttct ctcgagttta taatctccac cagaagaatg 1201 gcttcacatg tatgctcatt ggagagatgt ttgaactcat gcagttcctc tttgtggttg 1261 ccttcaccac cttcctggtt agctgtgtgg actatgacat cctctttgcc aacaagatgg 1321 tgaaccacag tctccatcct actgagcctg tcaaggtcac tctgccagat gcctttttgc 1381 ctgcccaagt ctgtagtgcc aggattcagg aaaatggctc tcttatcacc atcctggtca 1441 ttgctggtgt cttctggatc caccggctta tcaagtttat ctataacatt tgctgctatt 1501 gggagatcca ctccttctac ctacatgctc ttcggatccc aatgtctgcc cttccatact 1561 gcacatggca ggaagttcag gcccggattg tgcagacgca gaaagagcat cagatctgca 1621 tccacaagcg tgagctgaca gagttggaca tctaccatcg catcctacgt ttccagaact 1681 acatggtggc cctggtgaac aaatccctcc tgcctctgcg cttccgtctg cccggcctcg 1741 gagaggttgt cttcttcacc cgtggcctca agtacaactt tgagctcatc ctcttctggg 1801 gacccggctc tctgtttctc aatgagtgga gcctcaaggc cgagtacaaa cgtggagggc 1861 aacggctaga gctggcccag cgtctcagca accgcatctt gtggattggc atcgccaact 1921 tcctgctgtg tcccctcatc ctcatctggc agatcctcta tgccttcttc agctatgccg 1981 aggtgctgaa gagagagccg ggggccctgg gagcgcgttg ctggtcactc tatggccgtt 2041 gttacctccg ccacttcaat gagctggagc atgagctgca gtcccgcctc aaccgaggct 2101 acaagcccgc ctccaagtac atgaattgct tcttgtcacc gctgctgact ctgctggcca 2161 agaatggtgc cttcttcgct ggctctatcc tggctgtgct tattgccctc accatctatg 2221 atgaagatgt gttagctgtg gaacacgtcc tcaccacggt caccctcctg ggagtcacgg 2281 tgactgtgtg caggtccttc atcccagacc agcacatggt gttctgcccc gagcagctgc 2341 tccgagtgat tcttgcacac atccactaca tgcctgacca ctggcagggt aatgcccacc 2401 gctcgcagac ccgggacgag tttgcccagc tcttccagta caaggcagtg ttcatcttgg 2461 aggagttgct gagtcccatc gtcacacccc tcattctcat cttctgcctc cgccctcggg 2521 ccctggagat catagacttc ttccgcaact ttacggtcga ggtcgtgggt gtgggagaca 2581 cctgctcctt tgctcagatg gacgttcgcc agcatggcca tcctcagtgg ctgtctggag 2641 ggcagacaga ggcctcagtg taccagcaag ccgaggacgg gaagaccgag ttgtcgctca 2701 tgcactttgc catcaccaat cccggctggc agccccctcg tgagagcaca gctttcctgg 2761 gcttcctcaa ggagcaggtg cagcgagatg gagcagctgc tggcctggcc cagggtggcc 2821 tgctccccga gaatgccctc ttcacatcca tccagtcctt acagtctgag tccgagccac 2881 tgagccttat tgcaaatgtg gtagcaggct catcctgccg aggaccctca ctgtccagag 2941 acctgcaggg ctccaggcac agggctgatg ttgcttctgc ccttcgatcc ttctcccctc 3001 tgcagcctgg agcggcccct caaggccggg ttcccagtac catgacaggc tctggagtgg 3061 atgccaggac agccagctct gggagtagcg tgtgggaagg acagctgcag agcctggtgc 3121 tgtccgaata cgcgtccacc gagatgagcc tgcacgccct gtacatgcac cagctccaca 3181 agcagcagac ccaggctgag cccgagcggc atgtgtggca ccgccgggag agtgatgaga 3241 gtggagagag tgcccctgaa gaggggggag agggtgcccg ggccccccaa cccatccccc 3301 gctcggccag ctatccctgt gctacacccc ggcctggagc acctgagacc accgccctgc 3361 atgggggctt ccagaggcgc tacgggggca tcacagatcc tggcacagtg ccccgtggcc 3421 cctctcactt ctccaggctg cccctgggag gatgggcaga agatggccag ccagcatcaa 3481 ggcacccaga gccggtgcca gaggagggct cagaggatga actcccccct caagtgcaca 3541 aggtaagcgg accgacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc 3601 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc 3661 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg 3721 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta 3781 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg 3841 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa 3901 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa 3961 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc 4021 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc 4081 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg 4141 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac 4201 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg 4261 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg 4321 cggccagcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 4861 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 4921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 4981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc 5221 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg 5281 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc 5341 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg 5401 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt 5461 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg 5521 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa 5581 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc 5641 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 5701 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 5761 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcgatccgc cacacccagc 5821 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 5881 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 5941 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 6001 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 6061 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 6121 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 6181 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 6241 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 6301 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 6361 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 6421 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 6481 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc 6541 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg 6601 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat 6661 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga 6721 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 6781 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc 6841 tcaggactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 6901 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 6961 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 7021 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 7081 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 7141 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 7201 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 7261 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 7321 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 7381 atttaacaaa aatttaacgc gaatttt //