LOCUS       V030722                 8793 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030722
VERSION     V030722
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..3473
                     /label="DTHD1(NM_001170700)"
                     /note="DTHD1(NM_001170700)"
                     /gene="DTHD1"
     CDS             3480..3545
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3569..4157
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4223..4271
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4473..4901
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4915..5244
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5311..6105
                     /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    6281..6402
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6451..6467)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6475..6491
                     /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(6499..6529)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6544..6565
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6853..7441)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7612..8472)
                     /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(8473..8577)
                     /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 tatggaatac atgggctcag
      781 gtaaactggg ccaaatattg aagcagattt ggagacaaaa ccagatgcta aagcaggcac
      841 tcttgggtga tgacctttgt gagggggctg gtggggccac ttggatggcc actgtggtct
      901 ttctgggtca ggaactcagc agtgcccttc accagctgct ggagcacacc tcaggcaccc
      961 tgcgttccac ctgccagcag ctgcatgtgc tgcttgacaa agagaatcaa tgtgtctcga
     1021 gaaaagaaat cattactttc atagactgcc tcataaaaat aactgaacat ttggggagca
     1081 ctgctgcact tctaaagaaa gaagaaaagg agatttgtaa tttatgcggc atgcatgatg
     1141 aatgtactcc acagcagaca atgtcctcca ttcaagatac caaagcagca gacattgctg
     1201 caagagggga actaaatgtc atagaaacag ctactgtttc tcccacaaat ggagaggaaa
     1261 gtcattacac aaaccaggtc cagttagaaa aaaataaaac acatatgagt tcagcattag
     1321 tggaaaaaga aaacaataca tcactgaatg gacgtgtact ggggcaagaa gagtcacaga
     1381 ataaaatgtt cccagataat gcagaaaatg aagatgataa acaaatagaa cacatgactg
     1441 ttgagaacat aaatggcaac agggaagaga ctcatggcat aattcagaca acagagacag
     1501 aaattcaaga gacttcagaa agcccaagag aagagatgac cacatcctca ataatatgtg
     1561 atatctccaa gaaatatata aatagtactc ttcccaatga ttcagagaat ataaagcaca
     1621 agaataacat aatggaaaag gagtatcttg atgtgctgag tgatgttact ggcccccaag
     1681 tgtcttgtta tattacagca ccatcatatg ttctacaaca actagaatgc cggataataa
     1741 atcacatgag ttctttaata gtgggtgata atgaagagtt agttagcaac gtcataacta
     1801 ttgaatgctc agataaggaa aagagagttc catttccaat aggcattgca attccattta
     1861 ctgcacgtta cagaggaaat tacagagata tcatggtgaa agtgtgtgac ataaaccttc
     1921 aatcaagtta cctaaaccca aattcactag aaggaatgaa gggaggttat aaggggacct
     1981 gtgcttcagt aaaagtttac aaattgggta tcttttctgt tgtgtcttgt ttaaagaaag
     2041 agtcgttcac agtaacaaag aaaggcctcg ctcttaagtc aagcatggat tcccgaatat
     2101 ccttaaatta ccctccagga gtttttacct ctccagtgct ggtgcagtta aagatccaac
     2161 cagttgaccc agctctggtg gcacatttaa aagcacagca agatactttc tactcagtcc
     2221 aatccacaag ccctctgatt cacattcagc acccatcaac ttatcctttt cagaagccag
     2281 tcactttgtt tttaccttgt tctccatacc ttgataaaaa caaccttggt tctgagatag
     2341 atcataaaag aagagcaagt gccacaataa ataggattac accttcgtat ttcaaccgga
     2401 caaaaattgc ctccataaga aaacctagga aaaatgccag tgaatgtttg aaattactgg
     2461 gattcagaag ccaagacagt ggttggtgtg ggcttgatga tgttgtgaaa accatacaga
     2521 gcggcttggt atcagttgaa ttgtatgaac atttggagag gtttattgta cttcacctct
     2581 cttccaccat ggacaatagt catttggtta cttttgtgaa atctttagag gaagccatgc
     2641 tcagcaccac tgcctgcata gtactgtctc accagaagga caatccacat agaatagctg
     2701 ttttagtggt gccttccaaa gatttaagcc aggtgcttaa ggacctgcac ttggaagggt
     2761 ttggaggacc tccagagcca tctcgtcatt tccaagttcg agaaggagaa caacttcttt
     2821 taagatttac tggaaacata tttgcttcaa gcaacgggaa ggattatgga aaagactaca
     2881 cacttatttt tcacttgcaa agaaaaccta ggctggaact ccaaatcaaa gaagtggacg
     2941 aatttggaaa ctatagttgc cctcattaca aaggcaccat tgtcgtttat aaagtaccta
     3001 aaggaaagat agtccccaac ttgaatcaat ctctcgtaat taatgaaaac cattctcagt
     3061 tgccaatttg caaattacca ttgaaattgc caaagcataa gaaattaatc aaccgtccac
     3121 agagtaccaa aagagtttct aaggatcctg tagaagccct ttgggataac ttgctccatt
     3181 ggctggctga ggagctctca gaagaaaatg ctgagtctct ttcctcaact ctccctctgc
     3241 gccgtagcac cattcagctc atcaaactca agaaccctga tgatctcaca gaacagatcc
     3301 acgagtttct ttgcttctgg aaaaaatcgc ttccaacttt caccgacaaa cttcgcctcc
     3361 tggctcgaca tctccgcaag attggcagga gtgatcttgc agaagagctc aaattcaagt
     3421 gggaaaataa agtgttcact gaaccacagc agtgttttga tgtagcccct gagggatccg
     3481 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     3541 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3601 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3661 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3721 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3781 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3841 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3901 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3961 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4021 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4081 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4141 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     4201 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     4261 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4321 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4381 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     4441 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4501 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4561 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4621 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4681 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4741 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4801 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4861 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4921 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4981 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5041 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5101 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5161 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5221 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5281 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5341 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5401 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5461 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5521 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5581 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5641 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5701 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5761 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5821 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5881 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5941 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6001 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6061 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6121 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6181 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6241 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6301 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6361 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6421 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6601 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7021 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     7081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7261 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8521 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8581 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8641 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     8701 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     8761 ttgaccgaca attgcatgaa gaatctgctt agg
//
