LOCUS       V021342                 7652 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021342
VERSION     V021342
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..2278
                     /label="Nfia(NM_001122952)"
                     /note="Nfia(NM_001122952)"
                     /gene="Nfia"
     CDS             2318..2344
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2348..2374
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2378..2404
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2428..3016
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3082..3130
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3332..3760
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3774..4103
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4170..4964
                     /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    5140..5261
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5310..5326)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5334..5350
                     /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(5358..5388)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5403..5424
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5712..6300)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6471..7331)
                     /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(7332..7436)
                     /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 ccatgaagct tgctgacagc gtaatggcag ggaaagcttc
      721 cgacggctcc atcaaatggc agctttgcta cgacatctcg gccagaactt ggtggatgga
      781 tgagtttcat cctttcattg aagcacttct gccccatgtc cgcgccttcg cctacacatg
      841 gttcaacctg caggcccgaa agcggaaata cttcaaaaaa catgagaagc gcatgtcgaa
      901 agaagaggag agggccgtga aggatgaact gctaagcgag aagcccgagg tcaagcagaa
      961 gtgggcttcc cgacttctgg ccaagttacg gaaagatatc cgacccgagt accgagagga
     1021 ttttgttctt acagttacag ggaaaaaacc tccatgctgt gttctttcca accctgatca
     1081 gaaaggcaag atgcggagaa ttgactgcct ccgccaggca gataaagtat ggaggttgga
     1141 cctcgtcatg gtgatcttgt tcaaaggtat tccgctggaa agtactgatg gcgagcgcct
     1201 tgtaaagagt ccacagtgct ctaatccagg gctctgtgtc cagccccatc acataggggt
     1261 ttctgtaaag gaactcgatt tatatttggc atactttgta catgcagcag attcaagtca
     1321 atctgaaagt cccagccagc caagtgaagc tgacattaag gaccagccag aaaatggaca
     1381 tttgggcttc caggacagct ttgtcacatc aggtgttttc agtgtgactg agctagtaag
     1441 agtgtcacaa acaccaatag ctgcaggaac cggccccaat ttttctctct ctgatttgga
     1501 aagttcttca tactacagca tgagtccagg agcaatgagg aggtctctgc ccagcacatc
     1561 ctctaccagc tctacaaagc gcctcaagtc tgtggaggat gaaatggaca gtcctggtga
     1621 agaaccattt tacacaggcc aagggcgctc cccagggagt ggcagccagt ccagtggatg
     1681 gcatgaagta gagccaggct tgccatctcc aagcacactg aagaagtctg agaagtctgg
     1741 tttcagcagc ccctcccctt cgcagacctc ctccctggga acagcattca cacagcatca
     1801 ccgacctgtc attacaggac ccagagcaag tccacatgcg acgccatcga ctctccactt
     1861 tccaacgtca cccatcatcc agcagcctgg gccttacttc tcacacccag ccatccgtta
     1921 ccaccctcag gagacgctga aagagtttgt ccaacttgtc tgtcctgatg ctggtcagca
     1981 agctggacag gtggggttcc tcaatcccaa tggaagcagt caaggcaagg tgcacaaccc
     2041 attcctcccc accccaatgt tgccgccgcc gccaccacca ccgatggcca ggcctgtgcc
     2101 tctgcccatg ccagacacca agcctccaac cacatcaaca gaaggaggtg cagcctcccc
     2161 cacctcaccg acctactcga cacccagcac ctcccccgca aaccgattcg tcagtgttgg
     2221 accacgggat ccaagctttg taaatatccc tcaacagaca cagtcctggt acctgggaga
     2281 attctgcaga tatccagcac agtggcggcc gctcgagtac ccatacgacg tacctgatta
     2341 tgcaacttat ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta
     2401 cgcatagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     2461 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2521 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2581 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2641 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2701 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2761 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2821 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2881 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2941 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3001 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     3061 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     3121 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3181 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3241 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     3301 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3361 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3421 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3481 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3541 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3601 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3661 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     3721 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3781 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3901 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3961 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4021 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4081 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4141 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4201 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4261 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4321 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4381 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4441 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4501 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4561 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4621 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4681 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4741 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4801 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4861 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4921 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4981 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5041 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5101 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5161 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5221 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5281 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5881 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6121 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7441 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     7501 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     7561 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     7621 tgaccgacaa ttgcatgaag aatctgctta gg
//
