LOCUS       V023016                 8910 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023016
VERSION     V023016
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"
     regulatory      895..904
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             934..4398
                     /label="Ciita(NM_001302618)"
                     /note="Ciita(NM_001302618)"
                     /gene="Ciita"
     promoter        complement(4462..4480)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    4506..4730
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4776..5204
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5218..5548
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5615..6409
                     /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    6583..6704
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6753..6769)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6777..6793
                     /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(6801..6831)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6846..6867
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7155..7743)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7914..8774)
                     /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(8775..8879)
                     /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 gcttgccacc
      901 atggactaca aggacgacga tgataagggt accatgaacc acttccaggc catcctggcc
      961 caagtacaga cactgctctc cagccagaag cccaggcagg tgcgggccct cctggatggc
     1021 ctgctggaag aagagctgct ctcacgggaa taccactgtg ccttgctgca tgagcctgat
     1081 ggtgatgccc tggcccggaa gatttccctg accctgctgg agaaagggga cttagacttg
     1141 actttcttga gctgggtctg caacagtctg caggctccca cggtagagag gggcaccagc
     1201 tacagggacc atggagacca tagtctgtgt gccaccatgg atctgggatc tccagagggc
     1261 agctacctgg aactccttaa cagtgatgcc gaccccctac atctctacca cctctatgac
     1321 cagatggacc tggctgggga ggaggagatc gaactcagct cagagccaga cacagatacc
     1381 atcaactgcg accagttcag caagctgttg caggacatgg aactggatga agagacccgg
     1441 gaggcctatg ccaacattgc ggaactggat cagtacgtgt tccaggatac ccagctcgag
     1501 ggcctgagca aggacctctt catagagcac attggagcag aggaaggctt tggtgagaac
     1561 atagagatcc ctgtagaagc aggacagaag cctcagaaga gacgcttccc ggaagagcat
     1621 gctatggact caaagcacag gaagctagtg cccacctcta ggacctcact gaactatttg
     1681 gatctcccca ctgggcacat ccagatcttc accactctgc cccagggact ctggcaaatc
     1741 tcaggggctg gcacaggtct ctccagtgtc ctaatctacc acggtgagat gccccaggtc
     1801 aaccaagtgc tcccttcaag cagcctcagt atccccagtc tccccgagtc cccagaccgg
     1861 cctggctcca ccagcccctt cacaccatct gcagctgacc tgcccagcat gcccgaacct
     1921 gcgctgacct cccgtgtaaa tgagacagag gacacatctc cctccccatg ccaagagggt
     1981 cccgagtctt ccatcaagct tccaaaatgg ccagaggctg tggagcgatt ccagcactcc
     2041 ctacaggaca aatacaaggc attgccccag agcccaaggg gtcctctggt ggccgtggag
     2101 ctggtacggg ccaggctgga aagaggcagc aacaagagcc aggaaaggga gctggccact
     2161 cccgactgga cagagcgcca gctagcccac ggtggtctgg cagaggtact tcaggttgtc
     2221 agtgactgca ggcgaccagg agagacacag gtggtcgctg tgctgggcaa ggctggccag
     2281 ggaaagagcc actgggccag gacagtgagt cacacctggg catgtggcca gttgctacaa
     2341 tatgactttg tcttctatgt cccctgtcat tgcttggatc gtcccgggga cacctaccac
     2401 ctgcgggatc tgctctgtcc cccgagcctg cagccactgg ccatggatga cgaggtcctt
     2461 gattatatcg tgaggcagcc agaccgtgtt ctgctcatcc tagatgcttt cgaggagcta
     2521 gaggcccaag atggcctcct gcacggaccc tgtggatctc tgtccccaga gccctgctcc
     2581 ctccgaggac tgctggctgg gatcttccag cggaagctac tgcgaggctg cacactgctc
     2641 ctcacagccc ggccccgggg ccgcctggct cagagcctga gcaaggcaga tgccatcttt
     2701 gaggtgccca gcttctctac caagcaggcc aagacttaca tgaggcacta ctttgagaac
     2761 tcagggacag cggggaacca agacaaggcc ctgggcctcc tggagggcca gcctcttctc
     2821 tgcagctata gtcacagccc tgttgtgtgc agggctgtgt gccagctctc caaggccctg
     2881 ctagaacagg gcacagaggc ccagctacct tgtacactta caggactcta tgtcagcctg
     2941 ctaggtcctg cagctcagaa cagtcctccc ggagccttag tcgagctggc caagctggcc
     3001 tgggagctgg gacgaagaca ccaaagcacc ttgcaagaaa cccggttttc atccgtggag
     3061 gtgaaaacct gggcagtgac ccaaggcttg atgcagcaga ccctggagac cacggaggct
     3121 caactggcct tctccagttt tctgctacag tgtttcctgg gtgctgtgtg gctggcacag
     3181 tgcaatgaaa tcaaagacaa ggagctgcca cagtacctgg ccttgactcc gaggaagaag
     3241 agaccctatg acaactggct ggagggtgta ccacgctttc tggctggatt agttttccag
     3301 cctcgagccc actgcctggg agctctggtg gagcctgcag tggctgcagt ggcggatagg
     3361 aaacagaagg ttcttaccag gtacctgaag cgcctgaagc tggggacact ccgggcaggg
     3421 aggctgctgg agctgctcca ctgtgcccac gagacacagc aacctgggat atgggagcat
     3481 gttgcacacc agctccctgg ccacctctcc ttcctgggca cccggctcac acccccagat
     3541 gtgtatgtgc tgggcagggc cttggagaca gccagccagg acttctcctt ggaccttcgt
     3601 cagactggcg ttgagccttc tggactggga aacctcgtgg gactcagctg tgtcaccagt
     3661 ttcagggcct ccttgagtga tacaatggca ttatgggagt cccttcagca gcagggagaa
     3721 gcccagctac tccaggcggc agaggagaag ttcaccattg agccatttaa agccaaatcc
     3781 ccaaaggatg tggaagacct ggatcgtctc gtgcagaccc agaggctgag aaacccctca
     3841 gaagatgcag ccaaggatct tcctgccatc cgggacctta agaagctaga gtttgcgttg
     3901 ggccccatct tgggccccca ggctttcccc acactggcaa agatccttcc agccttctct
     3961 tctctgcaac acctggacct ggactcactt agtgagaaca agatcggaga caagggtgtg
     4021 tcgaagctct cagccacctt ccctcagctg aaggccctgg agacgctcaa cttgtcccaa
     4081 aacaacatca ctgatgtggg tgcctgcaag cttgcagaag ctctgccagc cctagccaag
     4141 tccctcctaa ggctgagctt gtacaataac tgcatctgtg acaaaggagc caagagcctg
     4201 gcacaagtac ttccggacat ggtgtccctg cgtgtgatgg atgtccagtt caacaagttc
     4261 acggctgccg gtgcccagca actggcctcc agccttcaga agtgccctca ggtggaaaca
     4321 ctggcaatgt ggacacccac tatccccttt ggggttcagg aacacctgca gcagctggat
     4381 gccaggatca gtctgagatg agaattctgc agatatccat cacactggcg gccgctcgag
     4441 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
     4501 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4561 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4621 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4681 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4741 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4801 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4861 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4921 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4981 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5041 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5101 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     5161 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     5221 tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat
     5281 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5341 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5401 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5461 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5521 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5581 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5641 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5701 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5761 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5821 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5881 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5941 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     6001 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6061 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     6121 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6181 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6241 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6301 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6361 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6421 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6481 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6541 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6601 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6661 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6721 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6781 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6841 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6901 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6961 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7021 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7081 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7141 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7201 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7261 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7321 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7381 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7441 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7501 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7561 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7621 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7681 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7741 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7801 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7861 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7921 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7981 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8041 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8101 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8161 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8221 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8281 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8341 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8401 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8461 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8521 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8581 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8641 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8701 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8761 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8821 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8881 acatttcccc gaaaagtgcc acctgacgtc
//
