LOCUS       V018387                 7835 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018387
VERSION     V018387
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             671..2515
                     /label="Nr1d1(NM_145434)"
                     /note="Nr1d1(NM_145434)"
                     /gene="Nr1d1"
     CDS             2522..2587
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2611..3199
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3265..3313
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3515..3943
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3957..4286
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4353..5147
                     /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    5323..5444
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5493..5509)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5517..5533
                     /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(5541..5571)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5586..5607
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5895..6483)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6654..7514)
                     /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(7515..7619)
                     /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 taccgccacc atgacgaccc tggactccaa taacaacaca ggtggtgtta tcacctacat
      721 tggctctagt ggctcctccc cgagccggac cagcccggag tccctctaca gtgacagctc
      781 caatggcagc ttccagtccc tgactcaagg ttgtcccaca tacttcccac catcacctac
      841 tggctccctc acccaggacc ctgcccgctc ttttggcagt gcgccaccca gtctcagtga
      901 tgatagctcc ccttcttctg catcatcgtc atcctcttca tcctcctcct ccttctataa
      961 cgggagcccc ccaggaagtc tacaagtggc catggaagac agcagccgag tgtcccccag
     1021 caagggcaca agcaacatta ccaagctgaa tggcatggtg ctactgtgta aggtgtgtgg
     1081 ggacgtggcc tcaggcttcc actatggagt gcacgcctgt gagggctgca agggcttttt
     1141 tcgccggagc atccaacaga atatccagta caaacggtgt ctgaaaaacg aaaactgctc
     1201 catcgttcgc atcaatcgca accgctgcca gcagtgtcgc ttcaagaagt gtctctccgt
     1261 tggcatgtct agagatgctg tgcgttttgg gcgcatcccc aagagagaga agcaacggat
     1321 gcttgccgag atgcagagcg ccatgaactt ggccaacaac caactgagca gcctgtgccc
     1381 tctagagacc tcacctactc cacatcccac ctctggctcc atgggcccct cacctcctcc
     1441 tgcaccagcc cccacacctt tggtgggctt ctctcagttc ccacaacagc tgacaccacc
     1501 caggtctccc agccctgagc ccaccatgga ggatgtgata tcccaggtcg cccgggccca
     1561 tcgagaaatc ttcacctatg cccatgacaa gttaggcacc tcacctggta acttcaatgc
     1621 caatcatgca tcaggtagcc cttcagctac aactccacac cgctgggaga gtcagggatg
     1681 cccgtctgcc cccaacgaca acaacctttt ggcggctcag cgtcataatg aagcgctgaa
     1741 tggtctacgc cagggccctt cctcctaccc gcctacctgg ccttctggcc ccacccacca
     1801 cagctgccac caacctaaca gcaatgggca ccgtctgtgc cccacccacg tatattcggc
     1861 cccagaaggg gaggcacctg ccaacagtct acggcaaggc aacaccaaga atgttctgct
     1921 ggcatgtccc atgaacatgt atccccatgg acgcagcggc cggactgtgc aggagatctg
     1981 ggaagacttc tctatgagct tcacgcctgc cgttcgggag gtggtagagt ttgccaaaca
     2041 catcccaggc ttccgtgacc tttctcagca cgaccaggtg accctgctta aggctggcac
     2101 ctttgaggtg ctgatggtgc gctttgcatc gttgttcaac gtgaaggacc agacagtgat
     2161 gttcctgagc cgcacaacct acagtctgca ggagctcggt gccatgggca tgggcgacct
     2221 gctcaatgcc atgtttgact tcagcgagaa gctcaactcc ctggcactta ccgaggagga
     2281 gctgggccta ttcaccgcgg tggtgcttgt ctctgcagac cgctcgggaa tggagaattc
     2341 cgcttcggtg gagcagctcc aggagacgct gctgcgggct cttcgggctc tggtgctgaa
     2401 gaaccggccc tcggagactt cccgcttcac caagctgctg ctcaagctgc cggacctgcg
     2461 gaccctgaac aacatgcatt ccgagaagct gctgtccttc cgggtggacg cccagctcga
     2521 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2581 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2761 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2821 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2881 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2941 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3001 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3061 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3121 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3181 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     3241 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     3301 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3361 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3421 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3481 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3541 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3601 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3661 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3721 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3781 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3841 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3901 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3961 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4141 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4201 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4261 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4321 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4381 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4441 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4501 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4561 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4621 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4681 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4741 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4801 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4861 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4921 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4981 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5041 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5101 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5161 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5221 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5281 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5341 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5401 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5461 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5521 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5581 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5641 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5701 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5761 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5821 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5881 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5941 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6001 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6061 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6121 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6181 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6241 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6301 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6361 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6421 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6481 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6541 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6601 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6661 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6721 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6781 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6841 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6901 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6961 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7021 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7081 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7141 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7201 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7261 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7321 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7381 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7441 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7501 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7561 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7621 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7681 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7741 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7801 gcttgaccga caattgcatg aagaatctgc ttagg
//
