LOCUS       V027870                 8981 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027870
VERSION     V027870
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"
     CDS             941..4471
                     /label="SREBF1(NM_001005291)"
                     /note="SREBF1(NM_001005291)"
                     /gene="SREBF1"
     CDS             4478..4543
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4581..4805
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4851..5279
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5293..5622
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5689..6483
                     /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    6657..6778
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6827..6843)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6851..6867
                     /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(6875..6905)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6920..6941
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7229..7814)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7985..8845)
                     /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(8846..8950)
                     /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 aagcttggta ccgagctcgg atccgccacc atggacgagc cacccttcag
      961 cgaggcggct ttggagcagg cgctgggcga gccgtgcgat ctggacgcgg cgctgctgac
     1021 cgacatcgaa ggtgaagtcg gcgcggggag gggtagggcc aacggcctgg acgccccaag
     1081 ggcgggcgca gatcgcggag ccatggattg cactttcgaa gacatgcttc agcttatcaa
     1141 caaccaagac agtgacttcc ctggcctatt tgacccaccc tatgctggga gtggggcagg
     1201 gggcacagac cctgccagcc ccgataccag ctccccaggc agcttgtctc cacctcctgc
     1261 cacattgagc tcctctcttg aagccttcct gagcgggccg caggcagcgc cctcacccct
     1321 gtcccctccc cagcctgcac ccactccatt gaagatgtac ccgtccatgc ccgctttctc
     1381 ccctgggcct ggtatcaagg aagagtcagt gccactgagc atcctgcaga cccccacccc
     1441 acagcccctg ccaggggccc tcctgccaca gagcttccca gccccagccc caccgcagtt
     1501 cagctccacc cctgtgttag gctaccccag ccctccggga ggcttctcta caggaagccc
     1561 tcccgggaac acccagcagc cgctgcctgg cctgccactg gcttccccgc caggggtccc
     1621 gcccgtctcc ttgcacaccc aggtccagag tgtggtcccc cagcagctac tgacagtcac
     1681 agctgccccc acggcagccc ctgtaacgac cactgtgacc tcgcagatcc agcaggtccc
     1741 ggtcctgctg cagccccact tcatcaaggc agactcgctg cttctgacag ccatgaagac
     1801 agacggagcc actgtgaagg cggcaggtct cagtcccctg gtctctggca ccactgtgca
     1861 gacagggcct ttgccgaccc tggtgagtgg cggaaccatc ttggcaacag tcccactggt
     1921 cgtagatgcg gagaagctgc ctatcaaccg gctcgcagct ggcagcaagg ccccggcctc
     1981 tgcccagagc cgtggagaga agcgcacagc ccacaacgcc attgagaagc gctaccgctc
     2041 ctccatcaat gacaaaatca ttgagctcaa ggatctggtg gtgggcactg aggcaaagct
     2101 gaataaatct gctgtcttgc gcaaggccat cgactacatt cgctttctgc aacacagcaa
     2161 ccagaaactc aagcaggaga acctaagtct gcgcactgct gtccacaaaa gcaaatctct
     2221 gaaggatctg gtgtcggcct gtggcagtgg agggaacaca gacgtgctca tggagggcgt
     2281 gaagactgag gtggaggaca cactgacccc acccccctcg gatgctggct cacctttcca
     2341 gagcagcccc ttgtcccttg gcagcagggg cagtggcagc ggtggcagtg gcagtgactc
     2401 ggagcctgac agcccagtct ttgaggacag caaggcaaag ccagagcagc ggccgtctct
     2461 gcacagccgg ggcatgctgg accgctcccg cctggccctg tgcacgctcg tcttcctctg
     2521 cctgtcctgc aaccccttgg cctccttgct gggggcccgg gggcttccca gcccctcaga
     2581 taccaccagc gtctaccata gccctgggcg caacgtgctg ggcaccgaga gcagagatgg
     2641 ccctggctgg gcccagtggc tgctgccccc agtggtctgg ctgctcaatg ggctgttggt
     2701 gctcgtctcc ttggtgcttc tctttgtcta cggtgagcca gtcacacggc cccactcagg
     2761 ccccgccgtg tacttctgga ggcatcgcaa gcaggctgac ctggacctgg cccggggaga
     2821 ctttgcccag gctgcccagc agctgtggct ggccctgcgg gcactgggcc ggcccctgcc
     2881 cacctcccac ctggacctgg cttgtagcct cctctggaac ctcatccgtc acctgctgca
     2941 gcgtctctgg gtgggccgct ggctggcagg ccgggcaggg ggcctgcagc aggactgtgc
     3001 tctgcgagtg gatgctagcg ccagcgcccg agacgcagcc ctggtctacc ataagctgca
     3061 ccagctgcac accatgggga agcacacagg cgggcacctc actgccacca acctggcgct
     3121 gagtgccctg aacctggcag agtgtgcagg ggatgccgtg tctgtggcga cgctggccga
     3181 gatctatgtg gcggctgcat tgagagtgaa gaccagtctc ccacgggcct tgcattttct
     3241 gacacgcttc ttcctgagca gtgcccgcca ggcctgcctg gcacagagtg gctcagtgcc
     3301 tcctgccatg cagtggctct gccaccccgt gggccaccgt ttcttcgtgg atggggactg
     3361 gtccgtgctc agtaccccat gggagagcct gtacagcttg gccgggaacc cagtggaccc
     3421 cctggcccag gtgactcagc tattccggga acatctctta gagcgagcac tgaactgtgt
     3481 gacccagccc aaccccagcc ctgggtcagc tgatggggac aaggaattct cggatgccct
     3541 cgggtacctg cagctgctga acagctgttc tgatgctgcg ggggctcctg cctacagctt
     3601 ctccatcagt tccagcatgg ccaccaccac cggcgtagac ccggtggcca agtggtgggc
     3661 ctctctgaca gctgtggtga tccactggct gcggcgggat gaggaggcgg ctgagcggct
     3721 gtgcccgctg gtggagcacc tgccccgggt gctgcaggag tctgagagac ccctgcccag
     3781 ggcagctctg cactccttca aggctgcccg ggccctgctg ggctgtgcca aggcagagtc
     3841 tggtccagcc agcctgacca tctgtgagaa ggccagtggg tacctgcagg acagcctggc
     3901 taccacacca gccagcagct ccattgacaa ggccgtgcag ctgttcctgt gtgacctgct
     3961 tcttgtggtg cgcaccagcc tgtggcggca gcagcagccc ccggccccgg ccccagcagc
     4021 ccagggcacc agcagcaggc cccaggcttc cgcccttgag ctgcgtggct tccaacggga
     4081 cctgagcagc ctgaggcggc tggcacagag cttccggccc gccatgcgga gggtgttcct
     4141 acatgaggcc acggcccggc tgatggcggg ggccagcccc acacggacac accagctcct
     4201 cgaccgcagt ctgaggcggc gggcaggccc cggtggcaaa ggaggcgcgg tggcggagct
     4261 ggagccgcgg cccacgcggc gggagcacgc ggaggccttg ctgctggcct cctgctacct
     4321 gccccccggc ttcctgtcgg cgcccgggca gcgcgtgggc atgctggctg aggcggcgcg
     4381 cacactcgag aagcttggcg atcgccggct gctgcacgac tgtcagcaga tgctcatgcg
     4441 cctgggcggt gggaccactg tcacttccag cctcgaggac tacaaagacc atgacggtga
     4501 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     4561 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4621 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4681 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4741 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4801 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4921 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4981 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5041 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5101 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5161 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5221 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5281 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5341 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5461 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5521 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5581 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5641 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5701 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5761 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5821 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5881 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5941 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6001 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6061 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6121 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6181 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6241 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6301 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6361 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6421 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6481 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6541 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6601 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6661 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6721 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6781 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6841 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6901 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7021 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7081 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7141 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7741 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8941 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
