LOCUS       V034058                 8786 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034058
VERSION     V034058
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..3442
                     /label="AR(NM_000044)"
                     /note="AR(NM_000044)"
                     /gene="AR"
     CDS             3449..3538
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3562..4150
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4216..4264
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4466..4894
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4908..5237
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5304..6098
                     /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    6274..6395
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6444..6460)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6468..6484
                     /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(6492..6522)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6537..6558
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6846..7434)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7605..8465)
                     /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(8466..8570)
                     /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 ccatggaagt gcagttaggg ctgggaaggg tctaccctcg
      721 gccgccgtcc aagacctacc gaggagcttt ccagaatctg ttccagagcg tgcgcgaagt
      781 gatccagaac ccgggcccca ggcacccaga ggccgcgagc gcagcacctc ccggcgccag
      841 tttgctgctg ctgcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
      901 gcagcagcag cagcagcagc aagagactag ccccaggcag cagcagcagc agcagggtga
      961 ggatggttct ccccaagccc atcgtagagg ccccacaggc tacctggtcc tggatgagga
     1021 acagcaacct tcacagccgc agtcggccct ggagtgccac cccgagagag gttgcgtccc
     1081 agagcctgga gccgccgtgg ccgccagcaa ggggctgccg cagcagctgc cagcacctcc
     1141 ggacgaggat gactcagctg ccccatccac gttgtccctg ctgggcccca ctttccccgg
     1201 cttaagcagc tgctccgctg accttaaaga catcctgagc gaggccagca ccatgcaact
     1261 ccttcagcaa cagcagcagg aagcagtatc cgaaggcagc agcagcggga gagcgaggga
     1321 ggcctcgggg gctcccactt cctccaagga caattactta gggggcactt cgaccatttc
     1381 tgacaacgcc aaggagttgt gtaaggcagt gtcggtgtcc atgggcctgg gtgtggaggc
     1441 gttggagcat ctgagtccag gggaacagct tcggggggat tgcatgtacg ccccactttt
     1501 gggagttcca cccgctgtgc gtcccactcc ttgtgcccca ttggccgaat gcaaaggttc
     1561 tctgctagac gacagcgcag gcaagagcac tgaagatact gctgagtatt cccctttcaa
     1621 gggaggttac accaaagggc tagaaggcga gagcctaggc tgctctggca gcgctgcagc
     1681 agggagctcc gggacacttg aactgccgtc taccctgtct ctctacaagt ccggagcact
     1741 ggacgaggca gctgcgtacc agagtcgcga ctactacaac tttccactgg ctctggccgg
     1801 accgccgccc cctccgccgc ctccccatcc ccacgctcgc atcaagctgg agaacccgct
     1861 ggactacggc agcgcctggg cggctgcggc ggcgcagtgc cgctatgggg acctggcgag
     1921 cctgcatggc gcgggtgcag cgggacccgg ttctgggtca ccctcagccg ccgcttcctc
     1981 atcctggcac actctcttca cagccgaaga aggccagttg tatggaccgt gtggtggtgg
     2041 tgggggtggt ggcggcggcg gcggcggcgg cggcggcggc ggcggcggcg gcggcggcgg
     2101 cgaggcggga gctgtagccc cctacggcta cactcggccc cctcaggggc tggcgggcca
     2161 ggaaagcgac ttcaccgcac ctgatgtgtg gtaccctggc ggcatggtga gcagagtgcc
     2221 ctatcccagt cccacttgtg tcaaaagcga aatgggcccc tggatggata gctactccgg
     2281 accttacggg gacatgcgtt tggagactgc cagggaccat gttttgccca ttgactatta
     2341 ctttccaccc cagaagacct gcctgatctg tggagatgaa gcttctgggt gtcactatgg
     2401 agctctcaca tgtggaagct gcaaggtctt cttcaaaaga gccgctgaag ggaaacagaa
     2461 gtacctgtgc gccagcagaa atgattgcac tattgataaa ttccgaagga aaaattgtcc
     2521 atcttgtcgt cttcggaaat gttatgaagc agggatgact ctgggagccc ggaagctgaa
     2581 gaaacttggt aatctgaaac tacaggagga aggagaggct tccagcacca ccagccccac
     2641 tgaggagaca acccagaagc tgacagtgtc acacattgaa ggctatgaat gtcagcccat
     2701 ctttctgaat gtcctggaag ccattgagcc aggtgtagtg tgtgctggac acgacaacaa
     2761 ccagcccgac tcctttgcag ccttgctctc tagcctcaat gaactgggag agagacagct
     2821 tgtacacgtg gtcaagtggg ccaaggcctt gcctggcttc cgcaacttac acgtggacga
     2881 ccagatggct gtcattcagt actcctggat ggggctcatg gtgtttgcca tgggctggcg
     2941 atccttcacc aatgtcaact ccaggatgct ctacttcgcc cctgatctgg ttttcaatga
     3001 gtaccgcatg cacaagtccc ggatgtacag ccagtgtgtc cgaatgaggc acctctctca
     3061 agagtttgga tggctccaaa tcacccccca ggaattcctg tgcatgaaag cactgctact
     3121 cttcagcatt attccagtgg atgggctgaa aaatcaaaaa ttctttgatg aacttcgaat
     3181 gaactacatc aaggaactcg atcgtatcat tgcatgcaaa agaaaaaatc ccacatcctg
     3241 ctcaagacgc ttctaccagc tcaccaagct cctggactcc gtgcagccta ttgcgagaga
     3301 gctgcatcag ttcacttttg acctgctaat caagtcacac atggtgagcg tggactttcc
     3361 ggaaatgatg gcagagatca tctctgtgca agtgcccaag atcctttctg ggaaagtcaa
     3421 gcccatctat ttccacaccc agctcgagga acaaaaacta atatcagagg aagatttgga
     3481 acagaaacta atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata
     3541 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     3601 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3661 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3721 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3781 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3841 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3901 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3961 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4021 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4081 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4141 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4201 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     4261 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4321 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4381 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     4441 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4621 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4801 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     4861 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4981 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5041 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5161 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5221 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5281 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5341 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5401 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5461 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5521 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5581 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5641 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5701 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5761 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5821 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5881 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5941 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6001 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6061 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     6121 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6181 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6241 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6301 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6361 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6421 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6481 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6541 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6601 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6961 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7021 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7261 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     7321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7501 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7561 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7621 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7681 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7741 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7801 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7861 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7921 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7981 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8041 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8101 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8161 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8221 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8281 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8341 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8401 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8461 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8521 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8581 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     8641 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     8701 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8761 acaattgcat gaagaatctg cttagg
//
