LOCUS       V027608                 8007 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027608
VERSION     V027608
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2687
                     /label="AR(NM_001348061)"
                     /note="AR(NM_001348061)"
                     /gene="AR"
     CDS             2694..2759
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2783..3371
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3437..3485
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3687..4115
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4129..4458
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4525..5319
                     /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    5495..5616
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5665..5681)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5689..5705
                     /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(5713..5743)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5758..5779
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6067..6655)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6826..7686)
                     /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(7687..7791)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga agtgcagtta gggctgggaa
      781 gggtctaccc tcggccgccg tccaagacct accgaggagc tttccagaat ctgttccaga
      841 gcgtgcgcga agtgatccag aacccgggcc ccaggcaccc agaggccgcg agcgcagcac
      901 ctcccggcgc cagtttgctg ctgctgcagc agcagcagca gcagcagcag cagcagcagc
      961 agcagcagca gcagcagcag cagcagcagc agcaagagac tagccccagg cagcagcagc
     1021 agcagcaggg tgaggatggt tctccccaag cccatcgtag aggccccaca ggctacctgg
     1081 tcctggatga ggaacagcaa ccttcacagc cgcagtcggc cctggagtgc caccccgaga
     1141 gaggttgcgt cccagagcct ggagccgccg tggccgccag caaggggctg ccgcagcagc
     1201 tgccagcacc tccggacgag gatgactcag ctgccccatc cacgttgtcc ctgctgggcc
     1261 ccactttccc cggcttaagc agctgctccg ctgaccttaa agacatcctg agcgaggcca
     1321 gcaccatgca actccttcag caacagcagc aggaagcagt atccgaaggc agcagcagcg
     1381 ggagagcgag ggaggcctcg ggggctccca cttcctccaa ggacaattac ttagggggca
     1441 cttcgaccat ttctgacaac gccaaggagt tgtgtaaggc agtgtcggtg tccatgggcc
     1501 tgggtgtgga ggcgttggag catctgagtc caggggaaca gcttcggggg gattgcatgt
     1561 acgccccact tttgggagtt ccacccgctg tgcgtcccac tccttgtgcc ccattggccg
     1621 aatgcaaagg ttctctgcta gacgacagcg caggcaagag cactgaagat actgctgagt
     1681 attccccttt caagggaggt tacaccaaag ggctagaagg cgagagccta ggctgctctg
     1741 gcagcgctgc agcagggagc tccgggacac ttgaactgcc gtctaccctg tctctctaca
     1801 agtccggagc actggacgag gcagctgcgt accagagtcg cgactactac aactttccac
     1861 tggctctggc cggaccgccg ccccctccgc cgcctcccca tccccacgct cgcatcaagc
     1921 tggagaaccc gctggactac ggcagcgcct gggcggctgc ggcggcgcag tgccgctatg
     1981 gggacctggc gagcctgcat ggcgcgggtg cagcgggacc cggttctggg tcaccctcag
     2041 ccgccgcttc ctcatcctgg cacactctct tcacagccga agaaggccag ttgtatggac
     2101 cgtgtggtgg tggtgggggt ggtggcggcg gcggcggcgg cggcggcggc ggcggcggcg
     2161 gcggcggcgg cggcgaggcg ggagctgtag ccccctacgg ctacactcgg ccccctcagg
     2221 ggctggcggg ccaggaaagc gacttcaccg cacctgatgt gtggtaccct ggcggcatgg
     2281 tgagcagagt gccctatccc agtcccactt gtgtcaaaag cgaaatgggc ccctggatgg
     2341 atagctactc cggaccttac ggggacatgc gtttggagac tgccagggac catgttttgc
     2401 ccattgacta ttactttcca ccccagaaga cctgcctgat ctgtggagat gaagcttctg
     2461 ggtgtcacta tggagctctc acatgtggaa gctgcaaggt cttcttcaaa agagccgctg
     2521 aagggaaaca gaagtacctg tgcgccagca gaaatgattg cactattgat aaattccgaa
     2581 ggaaaaattg tccatcttgt cgtcttcgga aatgttatga agcagggatg actctgggag
     2641 aaaaattccg ggttggcaat tgcaagcatc tcaaaatgac cagacccgga tccgactaca
     2701 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2761 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2821 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2881 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2941 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3001 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3061 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3121 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3181 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3241 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3301 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3361 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3421 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3481 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3541 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3601 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3661 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3841 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3901 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3961 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4021 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4081 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4141 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4201 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4321 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4381 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4441 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4501 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4561 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4621 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4681 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4741 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4801 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4861 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4921 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4981 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5041 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5101 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5161 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5221 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5281 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5341 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5401 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5461 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5581 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5641 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6961 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7141 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7201 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7261 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7321 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7381 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7441 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7501 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7561 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7621 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7681 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7741 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7801 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7861 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7921 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7981 gacaattgca tgaagaatct gcttagg
//
