LOCUS       V031027                 8271 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031027
VERSION     V031027
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..4418
                     /label="Srebf2(NM_033218)"
                     /note="Srebf2(NM_033218)"
                     /gene="Srebf2"
     CDS             4437..4466
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4485..4508
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4528..4544)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4555..5177
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5326..5914)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6243..6290
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6522..7316)
                     /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(7351..7708)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7710..7814)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7841..8269
                     /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 ggatgagagc agcgagctgg gcgttctgga gaccatggag accctcacgg
     1081 agctgggcga tgagctgact ctcggggaca tcgacgagat gctacagttt gtcagcaatc
     1141 aagtgggaga gttccctgat ttgttttcag agcaactttg tagctccttc cctggaggtg
     1201 gcagcaatgg tggcagtggc aacaacagca gtggcagagg caacaatggt ggagccacag
     1261 accctgcagt acagcggtca ttcagccagg tcccattatc taccttctct ccctcagctg
     1321 cctcccctca ggctccggcc ctgcaggtca aagtctctcc taccccacca agggcaaccc
     1381 ctgttctcca gccgcgaccc cagccccagc cccaacctcc agctcagctg cagcagcaga
     1441 cggtgatgat caccccgacg ttcagcaccg ctccgcagac gaggatcatc cagcagcctt
     1501 tgatatacca gaacgcagcg accagctttc aagtcctgca gcctcaagtg caaagcctcg
     1561 tgacgtcccc gcaggtgcag cccgtcacca tccagcagca ggtgcagacg gtacaggccc
     1621 agcgggtgtt gacgcagaca gccaatggca cactgcagac ccttgccccc gccacggtac
     1681 agacggtcgc tgcgccacag gtgcagcagg tcccggtcct ggtgcaacct cagatcatta
     1741 agacagattc ccttgtgttg accacgctga agacagatgg cagccctgtc atggctgcag
     1801 tccagaaccc ggccctcacg gccctcactg cccccatcca gacagccgcc cttcaagtgc
     1861 caaccctggt gggcagcaac gggaccattc tgaccacaat gcctgtgatg atggggcaag
     1921 agaaagttcc tatcaagcaa gtgcctggcg gcgtgaagca gctggatcct cccaaagaag
     1981 gagagaggcg gacaacacac aatatcattg aaaagcgcta ccggtcctcc atcaacgaca
     2041 aaatcataga gttgaaggac ttagtcatgg ggacagatgc caagatgcac aagtctggcg
     2101 ttctgaggaa ggccattgat tacatcaaat atctgcagca ggtcaatcac aagctgcgcc
     2161 aggagaacat ggtgctgaag ctggccaatc agaaaaacaa gctcctgaag ggcatcgacc
     2221 tgggcagtct ggtggacagt gatgtggact tgaaaattga tgactttaac cagaatgtcc
     2281 ttctgatgtc tccgccggcc tccgactccg ggtcccaggc cggcttctct ccctattcca
     2341 ttgactctga gccgggcagc cctctgctgg atgacgcaaa ggtcaaggat gaaccggact
     2401 ctcctcctgt ggcactgggc atggtggacc gctctcgaat cctcttatgt gtcctcacct
     2461 tcctgggcct ctcctttaac cccttgactt ccttgctgca gtggggaggg gcccacaaca
     2521 ctgaccagca cccatactca ggctcgggcc gcagtgtact gtcactggag tcaggtgctg
     2581 ggggctggtt tgactggatg gttccaactc tcctcctgtg gctggtaaat ggtgtgattg
     2641 tcttgagcgt ctttgtgaag ctgttggtcc acggggagcc ggtgattcgc ccacactcac
     2701 gcccctcagt caccttctgg agacaccgga agcaggcaga cctagacctc gccaaaggtg
     2761 atttcgcagc tgctgctgcc aacctacaaa cctgcttatc cgtgctgggc cgggcgctgc
     2821 ccacctcccg cctggacttg gcctgcagcc tctcctggaa tgtgatccgc tacagcctgc
     2881 agaagctgcg cctggtacgc tggttactca agaaggtctt ccagcgctgg cgggctactc
     2941 ccgccactgc agctggcttt gaggatgagg ctaagagcag cgcaagggat gcggcactgg
     3001 cctaccacag gctgcaccag ctgcacatca cagggaagct tccggcggga tctgcctgtt
     3061 cagatgtgca catggctctg tgtgctgtga acctggccga gtgtgcggag gagaaaatcc
     3121 taccaagcac actgattgag atccacctga cagcggccat ggggctcaag acccgctgtg
     3181 gaggcaagct gggcttcttg gctagctact tcttaaatcg tgcccagagt ctgtgtggcc
     3241 cagagcatag cactgtcccc gactccctgc gctggctatg ccatcccttg ggccagaagt
     3301 ttttcatgga gcgaagctgg tcgatcaagt cagcagccaa ggagagcctg tactgcgccc
     3361 agaggagccc agctgacccc attgcccagg tacaccaggc gttctgcaaa aacctgctag
     3421 aaagagcggt ggagtccttg gtgaaacctc aggccaagaa gaaggcaggc gaccaggaag
     3481 aagagagctg tgaattttcc agtgctctag agtatttgaa attacttcat tcttttgtgg
     3541 actctgtggg atttgtgacc tcaccattct ccagcagttc cgtgctcagg tcggcccttg
     3601 gccccgatgt catctgccgg tggtggacgt ccgcagtcac catggccatc agctggctcc
     3661 agggagacga tgcagctgtg cgctctcgtt ttactgaagt agagcgggtc cccaaggccc
     3721 tggaagtgac cgagagtccc ttggtgaagg ccgtcttcta tacctgtaga gccatgcatg
     3781 cctccctctc tgggaaagct gacgggcagc agaactcctt ctgtcactgt gagagggcca
     3841 gtggtcacct gtggagcagt ctcaacgtca gcggcactac ttctgatcct tcacttaacc
     3901 atgtgatcca gctgttcacc tgtgacctgc tactgtcgct gaggacagca ctctggcaaa
     3961 agcaggccag tgccagccag ctcctgggtg agacctacca tgcatccggc actgagctgg
     4021 ccggcttcca gcgggacctg ggcagccttc gaaggctggc ccatagcttc cgcccagcct
     4081 accgcaaggt gttcctgcat gaagccactg tgcgcctgat ggcaggagcc agccctaccc
     4141 gtacacacca gctgctggag catagcctac ggcggcgccc cactcagaac accaagcatg
     4201 gagaggtaga cacctggcct ggccaacgag agcgagctac tgccatcctg ctggcctgcc
     4261 gccatcttcc cctctctttc ctgtcttccc cgggccagcg ggcagtgctg ctggctgaag
     4321 ctgcccgcac cctggagaag gtgggtgacc gccgttcctg cagtgactgc cagcagatga
     4381 ttgtcaagct gggcggtggc actgccatcg ctgcctccac gcgtacgcgg ccgctcgagc
     4441 agaaactcat ctcagaagag gatctggcag caaatgatat cctggattac aaggatgacg
     4501 acgataaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg
     4561 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4621 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4681 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4741 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4801 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     4861 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     4921 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     4981 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     5041 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     5101 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     5161 tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc actcaaaggc
     5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6061 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     6121 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     6181 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     6241 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     6301 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa
     6361 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     6421 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     6481 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     6541 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     6601 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     6661 tgatagcgat ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     6721 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     6781 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     6841 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     6901 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     6961 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     7021 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     7081 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     7141 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     7201 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     7261 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     7321 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc
     7381 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7441 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7501 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     7561 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     7621 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     7681 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt
     7741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     7861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     7921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     8041 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     8101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     8161 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     8221 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t
//
