LOCUS       V020251                 6863 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020251
VERSION     V020251
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             941..2395
                     /label="NLE1(NM_018096)"
                     /note="NLE1(NM_018096)"
                     /gene="NLE1"
     CDS             2402..2425
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2463..2687
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2733..3161
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3175..3504
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3571..4365
                     /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    4539..4660
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4709..4725)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4733..4749
                     /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(4757..4787)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4802..4823
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5111..5696)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5867..6727)
                     /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(6728..6832)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggcggcag cagtgccgga
      961 cgaggcggtg gcgcgcgatg tgcagcggtt gctagtgcag ttccaggatg agggcgggca
     1021 gctgctgggt tccccgttcg acgtgcccgt ggacatcacc ccggacaggc tgcagctcgt
     1081 gtgcaacgcg ctactggccc aggaggatcc cctgccactg gctttctttg tccacgatgc
     1141 tgagatcgtc tcctcactgg ggaagacgtt ggagtcccag gcagtggaga cagagaaggt
     1201 cctagacatc atctaccagc cacaggctat cttcagagtc cgggctgtga ctcgctgcac
     1261 cagctccttg gagggtcaca gtgaggcagt catttctgtg gccttcagcc ctacgggaaa
     1321 gtacctggcc agtggctctg gagacaccac cgtgcgcttc tgggatctca gcacagagac
     1381 accacatttc acatgcaagg gacacagaca ctgggtcctt agtatatcct ggtctccaga
     1441 tggcaggaag ctggcctcag gctgcaagaa tggccagatt ctcctctggg acccaagcac
     1501 agggaagcag gtgggcagga ccctcgctgg ccacagcaag tggatcacag gcctgagctg
     1561 ggagcccctc catgcgaacc ctgagtgccg ctatgtggcc agcagctcca aggatggcag
     1621 tgtgcggatc tgggacacaa ctgcaggccg ctgtgagcgc atcctcaccg ggcacaccca
     1681 gtcggtcacc tgtctccggt ggggagggga cgggcttctc tactctgcct cccaggaccg
     1741 caccatcaaa gtctggagag ctcatgacgg tgtgctgtgc cggactctgc aaggccacgg
     1801 ccactgggtg aacaccatgg ccctcagcac tgactatgcc ctgcgcactg gggcctttga
     1861 acctgctgag gcctcagtta atccccaaga cctccaagga tccttgcagg agttgaagga
     1921 gagggctctg agccgataca acctcgtgcg gggccagggt ccagagaggc tggtgtctgg
     1981 ctccgacgac ttcaccttat tcctgtggtc cccagcagag gacaaaaagc ctctcactcg
     2041 gatgacagga caccaagctc tcatcaacca ggtgctcttc tctcctgact cccgcatcgt
     2101 ggctagtgcc tcctttgaca agtccatcaa gctgtgggat ggcaggacgg gcaagtacct
     2161 ggcttcccta cgcggccacg tggctgccgt gtaccagatt gcgtggtcag ctgacagtcg
     2221 gctcctggtc agcggcagca gtgacagcac actgaaggtg tgggatgtga aggcccagaa
     2281 gctggccatg gacctgcccg gccacgcgga tgaggtatat gctgttgact ggagtccaga
     2341 tggccagaga gtggcaagtg gtgggaagga caaatgcctc cggatatgga ggagactcga
     2401 ggattacaag gatgacgacg ataagtagtg agggcccgtt taaacccgct gatcagcctc
     2461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2641 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2701 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2761 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2821 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2881 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2941 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3001 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3061 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3121 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3181 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3301 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3361 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3421 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3481 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3541 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3601 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3661 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3721 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3781 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3841 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3901 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3961 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4021 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4081 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4141 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4201 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4261 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4321 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4381 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4441 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4501 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4561 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4621 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4681 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5641 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6181 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6481 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6841 cccgaaaagt gccacctgac gtc
//
