LOCUS       V031020                 6678 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031020
VERSION     V031020
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..2825
                     /label="Esr1(NM_007956)"
                     /note="Esr1(NM_007956)"
                     /gene="Esr1"
     CDS             2844..2873
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2892..2915
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2935..2951)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2962..3584
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4650..4697
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4929..5723)
                     /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(5758..6115)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6117..6221)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6248..6676
                     /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 gaccatgacc cttcacacca aagcctcggg aatggccttg ctgcaccaga
     1081 tccaagggaa cgagctggag cccctcaacc gcccgcagct caagatgccc atggagaggg
     1141 ccctgggcga ggtatacgtg gacaacagca agcccactgt gttcaactac cccgagggcg
     1201 ccgcctacga gttcaacgcc gccgccgccg ccgccgccgc cgcctcggcg ccggtctacg
     1261 gccagtcggg catcgcctac ggccccgggt cggaggcggc cgccttcagt gccaacagcc
     1321 tgggggcttt cccccagctc aacagcgtgt cgcctagccc gctgatgctg ctgcacccgc
     1381 cgccgcagct gtctcctttc ctgcacccgc acggccagca ggtgccctac tacctggaga
     1441 acgagcccag cgcctacgcc gtgcgcgaca ccggccctcc cgccttctac aggtctaatt
     1501 ctgacaatcg acgccagaat ggccgagaga gactgtccag cagtaacgag aaaggaaaca
     1561 tgatcatgga gtctgccaag gagactcgct actgtgccgt gtgcaatgac tatgcctctg
     1621 gctaccatta tggggtctgg tcctgcgaag gctgcaaggc tttctttaag agaagcattc
     1681 aaggacacaa tgactacatg tgtccagcta caaaccaatg caccattgac aagaaccgga
     1741 ggaagagttg ccaggcctgt cggctgcgca agtgttacga agtgggcatg atgaaaggcg
     1801 gcatacggaa agaccgccga ggagggagaa tgttgaagca caagcgtcag agagatgact
     1861 tggaaggccg aaatgaaatg ggtgcttcag gagacatgag ggctgccaac ctttggccaa
     1921 gccctcttgt gattaagcac actaagaaga atagccctgc cttgtccttg acagctgacc
     1981 agatggtcag tgccttgttg gatgctgaac cgcccatgat ctattctgaa tatgatcctt
     2041 ctagaccctt cagtgaagcc tcaatgatgg gcttattgac caacctagca gatagggagc
     2101 tggttcatat gatcaactgg gcaaagagag tgccaggctt tggggacttg aatctccatg
     2161 atcaggtcca ccttctcgag tgtgcctggc tggagattct gatgattggt ctcgtctggc
     2221 gctccatgga acacccgggg aagctcctgt ttgctcctaa cttgctcctg gacaggaatc
     2281 aaggtaaatg tgtggaaggc atggtggaga tctttgacat gttgctggct acgtcaagtc
     2341 ggttccgcat gatgaacctg cagggagaag agtttgtgtg cctcaaatcc atcattttgc
     2401 ttaattccgg agtgtacacg tttctgtcca gcaccttgaa gtctctggaa gagaaggacc
     2461 acatccaccg tgtcctggac aagatcacag acactttgat ccacctgatg gccaaagctg
     2521 gcctgactct gcagcagcag catcgccgcc tagctcagct ccttctcatt ctttcccata
     2581 tccggcacat gagtaacaaa ggcatggagc atctctacaa catgaaatgc aagaacgttg
     2641 tgcccctcta tgacctgctc ctggagatgt tggatgccca ccgccttcat gccccagcca
     2701 gtcgcatggg agtgccccca gaggagccca gccagaccca gctggccacc accagctcca
     2761 cttcagcaca ttccttacaa acctactaca tacccccgga agcagagggc ttccccaaca
     2821 cgatcacgcg tacgcggccg ctcgagcaga aactcatctc agaagaggat ctggcagcaa
     2881 atgatatcct ggattacaag gatgacgacg ataaggttta aacggccggc cgcggtcata
     2941 gctgtttcct gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     3001 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     3061 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     3121 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     3181 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     3241 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3301 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3361 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3421 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     3481 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     3541 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4441 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     4501 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     4561 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     4621 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     4681 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     4741 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     4801 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     4861 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     4921 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     4981 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     5041 atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag
     5101 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     5161 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     5221 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     5281 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     5341 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     5401 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     5461 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     5521 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     5581 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     5641 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     5701 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     5761 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5821 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5881 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5941 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6001 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     6061 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     6121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6241 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6301 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6661 aaatttaacg cgaatttt
//
