LOCUS       V022324                 8202 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022324
VERSION     V022324
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             898..924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             940..3690
                     /label="HK2(NM_000189)"
                     /note="HK2(NM_000189)"
                     /gene="HK2"
     promoter        complement(3754..3772)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    3798..4022
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4068..4496
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4510..4840
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4907..5701
                     /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    5875..5996
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6045..6061)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6069..6085
                     /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(6093..6123)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6138..6159
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6447..7035)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7206..8066)
                     /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(8067..8171)
                     /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 ggagacccgc caccatgtac
      901 ccatacgacg tcccagacta cgctggaagc ttggtaccga tgattgcctc gcatctgctt
      961 gcctacttct tcacggagct caaccatgac caagtgcaga aggttgacca gtatctctac
     1021 cacatgcgcc tctctgatga gaccctcttg gagatctcta agcggttccg caaggagatg
     1081 gagaaagggc ttggagccac cactcaccct actgcagcag tgaagatgct gcccaccttt
     1141 gtgaggtcca ctccagatgg gacagaacac ggagagttcc tggctctgga tcttggaggg
     1201 accaacttcc gtgtgctttg ggtgaaagta acggacaatg ggctccagaa ggtggagatg
     1261 gagaatcaga tctatgccat ccctgaggac atcatgcgag gcagtggcac ccagctgttt
     1321 gaccacattg ccgaatgcct ggctaacttc atggataagc tacaaatcaa agacaagaag
     1381 ctcccactgg gttttacctt ctcgttcccc tgccaccaga ctaaactaga cgagagtttc
     1441 ctggtctcat ggaccaaggg attcaagtcc agtggagtgg aaggcagaga cgttgtggct
     1501 ctgatccgga aggccatcca gaggagaggg gactttgata tcgacattgt ggctgtggtg
     1561 aatgacacag ttgggaccat gatgacctgt ggttatgatg accacaactg tgagattggt
     1621 ctcattgtgg gcacgggcag caacgcctgc tacatggaag agatgcgcca catcgacatg
     1681 gtggaaggcg atgaggggcg gatgtgtatc aatatggagt ggggggcctt cggggacgat
     1741 ggctcgctca acgacattcg cactgagttt gaccaggaga ttgacatggg ctcactgaac
     1801 ccgggaaagc aactgtttga gaagatgatc agtgggatgt acatggggga gctggtgagg
     1861 cttatcctgg tgaagatggc caaggaggag ctgctctttg gggggaagct cagcccagag
     1921 cttctcaaca ccggtcgctt tgagaccaaa gacatctcag acattgaagg ggagaaggat
     1981 ggcatccgga aggcccgtga ggtcctgatg cggttgggcc tggacccgac tcaggaggac
     2041 tgcgtggcca ctcaccggat ctgccagatc gtgtccacac gctccgccag cctgtgcgca
     2101 gccaccctgg ccgccgtgct gcagcgcatc aaggagaaca aaggcgagga gcggctgcgc
     2161 tctactattg gggtcgacgg ttccgtctac aagaaacacc cccattttgc caagcgtcta
     2221 cataagaccg tgcggcggct ggtgcccggc tgcgatgtcc gcttcctccg ctccgaggat
     2281 ggcagtggca aaggtgcagc catggtgaca gcagtggctt accggctggc cgatcaacac
     2341 cgtgcccgcc agaagacatt agagcatctg cagctgagcc atgaccagct gctggaggtc
     2401 aagaggagga tgaaggtaga aatggagcga ggtctgagca aggagactca tgccagtgcc
     2461 cccgtccgga tgctgcccac ctacgtgtgt gctaccccgg acggcacaga gaaaggggac
     2521 ttcttggcct tggaccttgg aggaacaaat ttccgggtcc tgctggtccg tgttcggaat
     2581 gggaagtggg gtggagtgga gatgcacaac aagatctacg ccatcccgca ggaggtcatg
     2641 cacggcaccg gggacgagct ctttgaccac attgtccagt gcatcgcgga cttcctcgag
     2701 tacatgggca tgaagggcgt gtccctgcct ctgggtttta ccttctcctt cccctgccag
     2761 cagaacagcc tggacgagag catcctcctc aagtggacaa aaggcttcaa ggcatctggc
     2821 tgcgagggcg aggacgtggt gaccctgctg aaggaagcga tccaccggcg agaggagttt
     2881 gacctggatg tggttgctgt ggtgaacgac acagtcggaa ctatgatgac ctgtggcttt
     2941 gaagaccctc actgtgaagt tggcctcatt gttggcacgg gcagcaatgc ctgctacatg
     3001 gaggagatgc gcaacgtgga actggtggaa ggagaagagg ggcggatgtg tgtgaacatg
     3061 gaatgggggg ccttcgggga caatggatgc ctagatgact tccgcacaga atttgatgtg
     3121 gctgtggatg agctttcact caaccccggc aagcagaggt tcgagaaaat gatcagtgga
     3181 atgtacctgg gtgagattgt ccgtaacatt ctcatcgatt tcaccaagcg tggactactc
     3241 ttccgaggcc gcatctcaga gcggctcaag acaaggggca tctttgaaac caagttcttg
     3301 tctcagattg agagtgactg cctggccctg ctgcaagtcc gagccatcct gcaacactta
     3361 gggcttgaga gcacctgtga cgacagcatc attgttaagg aggtgtgcac tgtggtggcc
     3421 cggcgggcag cccagctctg tggcgcaggc atggccgctg tggtggacag gatacgagaa
     3481 aaccgtgggc tggacgctct caaagtgaca gtgggtgtgg atgggaccct ctacaagcta
     3541 catcctcact ttgccaaagt catgcatgag acagtgaagg acctggctcc gaaatgtgat
     3601 gtgtctttcc tgcagtcaga ggatggcagc gggaaggggg cggcgctcat cactgctgtg
     3661 gcctgccgca tccgtgaggc tggacagcga taggaattct gcagatatcc atcacactgg
     3721 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     3781 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4141 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4201 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     4261 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     4441 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4501 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     4561 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4621 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4681 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4741 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4801 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4861 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4921 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4981 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5041 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5101 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5161 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5221 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5281 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5341 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5401 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5461 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5521 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5581 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5641 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5701 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5761 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5821 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5881 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5941 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6001 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6061 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6121 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6181 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6241 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6301 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6361 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6421 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6481 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6541 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6601 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6661 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6721 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6781 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6841 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6901 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6961 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7021 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7081 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7141 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7201 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7261 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7321 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7381 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7441 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7501 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7561 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7621 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7681 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7741 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7801 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7861 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7921 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7981 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8041 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8101 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8161 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
