LOCUS       V019315                 9404 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019315
VERSION     V019315
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..4084
                     /label="Srebf1(NM_001276707)"
                     /note="Srebf1(NM_001276707)"
                     /gene="Srebf1"
     CDS             4091..4156
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4180..4768
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4834..4882
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5084..5512
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5526..5855
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5922..6716
                     /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    6892..7013
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7062..7078)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7086..7102
                     /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(7110..7140)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7155..7176
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7464..8052)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8223..9083)
                     /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(9084..9188)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggacga gctacccttc ggtgaggcgg ctctggaaca
      721 ggcactggcc gaggtgtgcg aaatggacgc ggcgctcttg accgacatcg aagacatgct
      781 tcagctcatc aacaaccaag acagtgactt ccctggccta tttgatgccc cctatgctgg
      841 gggtgagaca ggagacacag gccccagcag ccctggtgcc agctctcctg agagcttctc
      901 ttctcctgct tctctgggct cctctctgga agccttcctg ggaggaccca aggtgacacc
      961 tgcacccttg tcccctccac catcggcacc cactgctgta aagatgtacc cgtccgtgcc
     1021 ccccttctcc cctgggcctg gaatcaaaga ggagccagtg ccactcacca tcctgcagcc
     1081 cccagcacca cagccatcgc cagggaccct gttgcctccg agcttccctc ctccacctgt
     1141 gcagctcagc cctgctcctg tgctggggta ctcaagcctg ccttccggct tctcaggaac
     1201 ccttcctggg aacacccagc agacgccatc tagcctgcca ctgggctcca cgccaggaat
     1261 ctcgcccacc cccttacaca cccaggtcca gagctcggcc gcccagcagc cgccgccagc
     1321 ctcagcagcc cctagaatga gcactgtggc ctcacagatc cagcaggtcc ccgttgtact
     1381 gcagccacac ttcatcaagg cagactcgct gctgctgaca gctgtaaaga cagacacagg
     1441 agccacaatg aagaccgcag gcatcaacac cctggctcct ggcacagccg tgcaggcagg
     1501 ccccttgcag accctggtga gtggagggac catcctggcc acagtaccac tggttgtgga
     1561 cacagacaaa ctgcccatcc accgactagc agctggtggc aaggccctgg gctcagctca
     1621 gagccgtggt gagaagcgca cagcccacaa tgccattgag aagcgctacc gttcctctat
     1681 caatgacaag attgtggagc tcaaggacct ggtggtgggc actgaggcaa agctgaataa
     1741 atctgctgtc ttgcgcaagg ccatcgacta catccgcttc ttacagcaca gcaaccagaa
     1801 actcaagcag gagaacctga ccctgcgaag tgctcacaaa agcaaatcac tgaaagacct
     1861 ggtgtcagct tgtggcagtg gaggaggcac agatgtgtct atggagggca tgaaacctga
     1921 agtggtagaa acgctgaccc ctccaccctc agacgccggc tcaccctccc agagtagccc
     1981 cttgtccttg ggcagcagag gcagcagcag tggtggcagt gactctgagc ccgacagccc
     2041 agcctttgag gataaccagg tgaaagccca gcggctgcct tcacatagcc gaggcatgct
     2101 ggaccgctcc cgcctggccc tgtgtgtact ggtcttcctg tgtctgacct gcaacccatt
     2161 ggcctcactg tttggctggg gcatcctcac tccctctgat gcttcgggtg tgcaccgtag
     2221 ttctgggcgc agcatgctgg aggccgagag cagagatggc tctaattgga cccagtggtt
     2281 gctgccaccc ctagtctggc tggccaatgg actactagtg ttggcctgct tggctcttct
     2341 ctttgtctac ggggaacctg tgaccaggcc acactccggc ccggctgtac acttctggag
     2401 acatcgcaaa caagctgacc tggatttggc ccggggagat tttgcccagg ccgctcaaca
     2461 gctgtggctg gccttgcaag ccctgggccg gcccctgccc acctcaaacc tggatctggc
     2521 ctgcagcctg ctttggaacc tcgtccgcca cctgctgcag cgtctttggg tgggccgctg
     2581 gctggcaggc caggctgggg gcctgcagag ggactacagg ctgagaaagg atgctcgtgc
     2641 cagtgcccga gatgcggctg tcgtctacca taagctgcac cagctgcatg ccatgggcaa
     2701 gtacacagga ggccatcttg ttgcttctaa cctggcactg agtgccctta acctggctga
     2761 gtgtgcagga gatgctatat ccatggcaac actggcagag atctacgtgg cagctgccct
     2821 aagggtcaaa accagcctcc ccagagcctt gcacttcttg acacgtttct tcctgagtag
     2881 tgcccgccag gcctgcctgg cacagagtgg tgcagtgcct cttgccatgc agtggctctg
     2941 ccaccctgta ggtcaccgtt tcttcgtgga tggggactgg gctgtacacg gtgcccccca
     3001 ggagagtctg tacagcgtgg ctgggaaccc agtggatcca ctggcccagg tgacccgact
     3061 attctgtgaa catctcctgg agcgagcatt gaactgtatc gctcagccca gcccaggggc
     3121 agctgatgga gacagggagt tctcagatgc tcttggatat ctacagttgc taaatagctg
     3181 ttctgacgct gtcggagctc ctgcgtgcag cttctctgtc agttccagca tggctaccac
     3241 cactggcaca gacccagtgg ccaagtggtg ggcctcactg acagccgtgg tgatccactg
     3301 gctgaggcgg gatgaggagg cagctgaacg cttataccca ctggtagagc acattcccca
     3361 agtgctgcag gaaactgaga gaccccttcc cagggcagct ctgtactcct tcaaggctgc
     3421 ccgggctctg ctggaccaca gaaaggtgga atccagccca gccagcctgg ccatctgtga
     3481 gaaggccagt gggtacctgc gggacagctt agcctctaca tcaactgcca gttccattga
     3541 caaggccatg cagctgctcc tgtgtgatct acttcttgtg gcccgcacca gcctatggcg
     3601 gcgccaacag tcagcagctt cagcccaggg agctcacggt accagcaatg gaccccaggc
     3661 ctctgctctg gagctgcgtg gtttccaaca tgacctgagc agcctgaggc gcttggcaca
     3721 gagcttccgg cctgctatga ggagggtctt cctacatgag gccacagctc ggctgatggc
     3781 aggagcaagt cctgcccgga cacaccagct cctggaccgc agtctgcgga ggcgggcagg
     3841 ttccagtggc aaaggaggcg ctgcagctga gctggagcct cgacccacat ggcgggagca
     3901 cacagaggcc ttgctgttgg cctcctgcta tctgccccct gccttcctgt cggcccccgg
     3961 gcagcgagtg agcatgctgg ctgaggcagc gcgcaccgtg gagaagcttg gcgatcaccg
     4021 gctcctgctt gactgccagc agatgctcct gcgcctgggt ggcgggacca ctgtcacttc
     4081 cagcctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     4141 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     4201 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4261 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4321 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4381 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4441 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4501 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4561 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4621 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4681 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4741 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     4801 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4861 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4921 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4981 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5041 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5101 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5161 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5221 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     5281 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5341 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5401 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     5461 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5521 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5581 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5641 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5701 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5761 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5821 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5881 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5941 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6001 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6061 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     6121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6421 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6481 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6601 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6721 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6781 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6841 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6901 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6961 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7021 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7081 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7141 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7201 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7261 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7321 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7381 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7441 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7501 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7561 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7621 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7681 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7741 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7801 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7861 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7921 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7981 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8041 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8101 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8161 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8221 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8281 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8341 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8401 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8461 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8521 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8581 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8641 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8701 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8761 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8821 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8881 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8941 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9001 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9061 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9121 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9181 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     9241 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9301 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9361 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
