LOCUS       V035339                 6638 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035339
VERSION     V035339
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             955..2538
                     /label="TH(NM_199292)"
                     /note="TH(NM_199292)"
                     /gene="TH"
     CDS             2560..2589
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2605..2622
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2651..2875
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2921..3349
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3363..3693
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        3741..3788
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             3807..4181
                     /codon_start=1
                     /gene="Sh&nbsp;ble from&nbsp;Streptoalloteichus
                     hindustanus"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin™"
                     /product="antibiotic-binding protein"
                     /transl_table=1
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD*"
                     /label="BleoR"
     polyA_signal    4311..4432
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4481..4497)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4505..4521
                     /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(4529..4559)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4574..4595
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4883..5471)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5642..6502)
                     /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(6503..6607)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatgccc
      961 acccccgacg ccaccacgcc acaggccaag ggcttccgca gggccgtgtc tgagctggac
     1021 gccaagcagg cagaggccat catggtaaga gggcagggcg ccccggggcc cagcctcaca
     1081 ggctctccgt ggcctggaac tgcagcccca gctgcatcct acacccccac cccaaggtcc
     1141 ccgcggttca ttgggcgcag gcagagcctc atcgaggacg cccgcaagga gcgggaggcg
     1201 gcggtggcag cagcggccgc tgcagtcccc tcggagcccg gggaccccct ggaggctgtg
     1261 gcctttgagg agaaggaggg gaaggccgtg ctaaacctgc tcttctcccc gagggccacc
     1321 aagccctcgg cgctgtcccg agctgtgaag gtgtttgaga cgtttgaagc caaaatccac
     1381 catctagaga cccggcccgc ccagaggccg cgagctgggg gcccccacct ggagtacttc
     1441 gtgcgcctcg aggtgcgccg aggggacctg gccgccctgc tcagtggtgt gcgccaggtg
     1501 tcagaggacg tgcgcagccc cgcggggccc aaggtcccct ggttcccaag aaaagtgtca
     1561 gagctggaca agtgtcatca cctggtcacc aagttcgacc ctgacctgga cttggaccac
     1621 ccgggcttct cggaccaggt gtaccgccag cgcaggaagc tgattgctga gatcgccttc
     1681 cagtacaggc acggcgaccc gattccccgt gtggagtaca ccgccgagga gattgccacc
     1741 tggaaggagg tctacaccac gctgaagggc ctctacgcca cgcacgcctg cggggagcac
     1801 ctggaggcct ttgctttgct ggagcgcttc agcggctacc gggaagacaa tatcccccag
     1861 ctggaggacg tctcccgctt cctgaaggag cgcacgggct tccagctgcg gcctgtggcc
     1921 ggcctgctgt ccgcccggga cttcctggcc agcctggcct tccgcgtgtt ccagtgcacc
     1981 cagtatatcc gccacgcgtc ctcgcccatg cactcccctg agccggactg ctgccacgag
     2041 ctgctggggc acgtgcccat gctggccgac cgcaccttcg cgcagttctc gcaggacatt
     2101 ggcctggcgt ccctgggggc ctcggatgag gaaattgaga agctgtccac gctgtactgg
     2161 ttcacggtgg agttcgggct gtgtaagcag aacggggagg tgaaggccta tggtgccggg
     2221 ctgctgtcct cctacgggga gctcctgcac tgcctgtctg aggagcctga gattcgggcc
     2281 ttcgaccctg aggctgcggc cgtgcagccc taccaagacc agacgtacca gtcagtctac
     2341 ttcgtgtctg agagcttcag tgacgccaag gacaagctca ggagctatgc ctcacgcatc
     2401 cagcgcccct tctccgtgaa gttcgacccg tacacgctgg ccatcgacgt gctggacagc
     2461 ccccaggccg tgcggcgctc cctggagggt gtccaggatg agctggacac ccttgcccat
     2521 gcgctgagtg ccattggcct cgagtctaga gggcccttcg aacaaaaact catctcagaa
     2581 gaggatctga atatgcatac cggtcatcat caccatcacc attgagttta aacccgctga
     2641 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2701 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2761 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2821 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2881 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3061 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3121 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3241 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     3301 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3361 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa
     3421 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3481 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3541 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3601 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3661 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3721 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3781 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     3841 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     3901 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     3961 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     4021 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     4081 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     4141 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     4201 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4261 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4381 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4441 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4501 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4561 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4861 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4921 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4981 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5041 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5101 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5221 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5281 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5341 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5401 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5461 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5521 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5581 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5641 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5701 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5761 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5821 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5881 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5941 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6001 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6061 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6301 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6481 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6541 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6601 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
