LOCUS       V042557                 8803 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042557
VERSION     V042557
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..4320
                     /label="Srebf1(NM_001358314)"
                     /note="Srebf1(NM_001358314)"
                     /gene="Srebf1"
     polyA_signal    4400..4624
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4670..5098
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5112..5441
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5508..6302
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6476..6597
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6646..6662)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6670..6686
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6694..6724)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6739..6760
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7048..7636)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7807..8667)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8668..8772)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggattgca catttgaaga catgctccag
     1021 ctcatcaaca accaagacag tgacttcccg ggcctgtttg acgcccccta tgctgggggt
     1081 gagacagggg acacaggccc cagcagccca ggtgccaact ctcctgagag cttctcttct
     1141 gcttctctgg cctcctctct ggaagccttc ctgggaggac ccaaggtgac acctgcaccc
     1201 ttgtcccctc caccatcggc acccgctgct ttaaagatgt acccgtccgt gtcccccttt
     1261 tcccctgggc ctgggatcaa agaggagcca gtgccactca ccatcctaca gcctgcagcg
     1321 ccacagccgt caccggggac cctcctgcct ccgagcttcc ccgcaccacc cgtacagctc
     1381 agccctgcgc ccgtgctggg ttactcgagc ctgccttcag gcttctcagg gacccttcca
     1441 ggaaacactc agcagccacc atctagcctg ccgctggccc ctgcaccagg agtcttgccc
     1501 acccctgccc tgcacaccca ggtccaaagc ttggcctccc agcagccgct gccagcctca
     1561 gcagccccta gaacaaacac tgtgacctca caggtccagc aggtcccagt tgtactgcag
     1621 ccacacttca tcaaggcaga ctcactgctg ctgacagctg tgaagacaga tgcaggagcc
     1681 accgtgaaga ctgcaggcat cagcaccctg gctcctggca cagccgtgca ggcaggtccc
     1741 ctgcagaccc tggtgagtgg agggaccatc ttggccacag tacctttggt tgtggacaca
     1801 gacaaactgc ccatccaccg actcgcagct ggcagcaagg ccctaggctc agctcagagc
     1861 cgtggtgaga agcgcacagc ccacaatgcc attgagaagc gctaccggtc ttctatcaat
     1921 gacaagattg tggagctcaa agacctggtg gtgggcactg aagcaaagct gaataaatct
     1981 gctgtcttgc gcaaggccat cgactacatc cgcttcttgc agcacagcaa ccagaagctc
     2041 aagcaggaga acctgaccct acgaagtgca cacaaaagca aatcactgaa ggacctggtg
     2101 tcagcttgtg gcagtggagg aggcacagat gtgtctatgg agggcatgaa acccgaagtg
     2161 gtggagacgc ttacccctcc accctcagac gccggctcac cctcccagag tagccccttg
     2221 tcttttggca gcagagctag cagcagtggt ggtagtgact ctgagcccga cagtccagcc
     2281 tttgaggata gccaggtcaa agcccagcgg ctgccttcac acagccgagg catgctggac
     2341 cgctcccgcc tggccctgtg tgtactggcc tttctgtgtc tgacctgcaa tcctttggcc
     2401 tcgcttttcg gctggggcat tctcactccc tctgatgcta cgggtacaca ccgtagttct
     2461 gggcgcagca tgctggaggc agagagcaga gatggctcta attggaccca gtggttgctg
     2521 ccacccctag tctggctggc caatggacta ctagtgttgg cctgcttggc tcttctcttt
     2581 gtctatgggg aacctgtgac taggccacac tctggcccgg ctgtacactt ctggagacat
     2641 cgcaaacaag ctgacctgga tttggcccgg ggagatttcc cccaggctgc tcaacagctg
     2701 tggctggccc tgcaagcgct gggccggccc ctgcccacct caaacctgga tctggcctgc
     2761 agtctgcttt ggaacctcat ccgccacctg ctccagcgtc tctgggtggg ccgctggctg
     2821 gcaggccagg ccgggggcct gctgagggac cgtgggctga ggaaggatgc ccgtgccagt
     2881 gcccgggatg cggctgttgt ctaccataag ctgcaccagc tgcatgccat gggcaagtac
     2941 acaggaggac atcttgctgc ttctaacctg gcactaagtg ccctcaacct ggctgagtgc
     3001 gcaggagatg ctatctccat ggcaacactg gcagagatct atgtggcagc ggccctgagg
     3061 gtcaaaacca gcctcccaag agccctgcac ttcttgacac gtttcttcct gagcagcgcc
     3121 cgccaggcct gcctagcaca gagcggctcg gtgcctcttg ccatgcagtg gctctgccac
     3181 cctgtaggtc accgtttctt tgtggacggg gactgggccg tgcacggtgc ccccccggag
     3241 agcctgtaca gcgtggctgg gaacccagtg gatccgctgg cccaggtgac ccggctattc
     3301 cgtgaacatc tcctagagcg agcgttgaac tgtattgctc agcccagccc aggggcagct
     3361 gacggagaca gggagttctc agatgccctt ggatatctgc agttgctaaa tagctgttct
     3421 gatgctgccg gggctcctgc ttgcagtttc tctgtcagct ccagcatggc tgccaccact
     3481 ggcccagacc cagtggccaa gtggtgggcc tcactgacag ctgtggtgat ccactggctg
     3541 aggcgggatg aagaggcagc tgagcgcttg tacccactgg tagagcatat cccccaggtg
     3601 ctgcaggaca ctgagagacc cctgcccagg gcagctctgt actccttcaa ggctgcccgg
     3661 gctctgctgg accacagaaa ggtggaatct agcccagcca gcctggccat ctgtgagaag
     3721 gccagtgggt acctgcggga cagcttagcc tctacaccaa ctggcagttc cattgacaag
     3781 gccatgcagc tgctcctgtg tgatctactt cttgtggccc gtaccagtct gtggcagcgg
     3841 cagcagtcac cagcttcagt ccaggtagct cacggtacca gcaatggacc ccaggcctct
     3901 gctctggagc tgcgtggttt ccaacatgac ctgagcagcc tgcggcggtt ggcacagagc
     3961 ttccggcctg ctatgaggag ggtattccta catgaggcca cagctcggct gatggcagga
     4021 gcaagtcctg cccggacaca ccagctcctg gatcgcagtc tgaggaggag ggcaggttcc
     4081 agtggcaaag gaggcactac agctgagctg gagccacggc ccacatggcg ggagcacacc
     4141 gaggccctgc tgttggcatc ctgctatctg ccccctgcct tcctgtcggc tcctgggcag
     4201 cgaatgagca tgctggccga ggcggcacgc accgtagaga agcttggcga tcaccggcta
     4261 ctgctggact gccagcagat gctcctgcgc ctgggcggcg gaaccaccgt cacttccagc
     4321 taggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     4381 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4621 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5041 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     5101 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5161 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5221 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5281 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5341 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5401 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5461 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5521 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5581 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5641 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5701 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5761 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5821 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5881 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5941 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6001 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     6061 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6121 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6181 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6241 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6301 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6361 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6421 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6601 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
