LOCUS       V018736                 8902 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018736
VERSION     V018736
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             907..3594
                     /label="N4BP1(NM_153029)"
                     /note="N4BP1(NM_153029)"
                     /gene="N4BP1"
     CDS             3607..3672
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             3682..3738
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3751..4470
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4502..4726
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4772..5200
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5214..5543
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5610..6404
                     /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    6578..6699
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6748..6764)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6772..6788
                     /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(6796..6826)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6841..6862
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7150..7735)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7906..8766)
                     /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(8767..8871)
                     /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 gccaccatgg cggcccgggc ggtgctggac gagttcactg cgccagctga gaaggcggag
      961 ctgctggagc agagccgcgg ccgtatcgag ggcctgtttg gcgtgagcct agccgtgctc
     1021 ggcgcgctag gggctgagga gccgctgccc gcgcgcatct ggctgcagct ctgcggggcg
     1081 caggaggcgg tgcacagcgc caaggaatat attaaaggaa tctgtgaacc tgaactagaa
     1141 gaaagagaat gttaccccaa ggacatgcac tgcatttttg ttggggcaga gagcctgttt
     1201 ctgaaaagct tgattcagga tacttgtgct gacctctgca ttctggacat tggccttctt
     1261 ggcatcagag gaagtgctga ggctgtggtc atggctagga gtcacattca acaatttgta
     1321 aagctctttg aaaataaaga gaacctaccc agtagtcaga aagaatcaga ggtgaaaagg
     1381 gaattcaaac aatttgttga agcccatgca gacaattaca caatggattt gttgattttg
     1441 cccacttcct tgaaaaaaga acttttgaca ctcacacaag gtgaggagaa tctctttgaa
     1501 acaggagatg atgaggttat tgaaatgaga gattctcaac aaacagagtt tacacagaat
     1561 gctgccacag ggctgaatat ttctagagat gaaactgttt tgcaggaaga ggcaagaaat
     1621 aaagctggga ctcctgtttc tgagcttaca aaacaaatgg acacagtcct ttctagctca
     1681 ccagatgtgc tttttgatcc aataaatggt ctaaccccag atgaagaggc actttccaat
     1741 gagagaattt gtcagaaaag gagattttct gattctgaag aaaggcatac gaagaagcag
     1801 ttttctttgg aaaatgttca ggagggggag attttacacg atgctaagac attggctgga
     1861 aatgtaatag ctgacctatc tgattcttct gctgattctg aaaatttaag tccagatata
     1921 aaagaaacta ctgaggaaat ggaatacaac atcctcgtaa acttttttaa aaccatgggc
     1981 tactcccaag aaattgttga aaaggtcatt aaggtgtatg gaccatctac tgaaccatta
     2041 ttgctcttag aggaaattga aaaagaaaat aaaagattcc aagaagacag agaattttca
     2101 gctggtacag tgtatccaga gaccaacaaa accaaaaata aaggtgttta tagcagcaca
     2161 aatgagctta caactgattc cactccaaag aaaacacagg ctcacacaca gcaaaatatg
     2221 gtagaaaaat tttctcagtt accattcaaa gtggaagcta aaccatgtac ctcaaattgt
     2281 agaattaata ctttcagaac agtgccaata gaacagaaac atgaagtctg gggttcaaac
     2341 cagaactaca tttgtaacac agaccctgaa actgatggcc tttcaccttc tgttgcctct
     2401 ccaagtccca aagaagtcaa ttttgtttca aggggagctt caagtcacca gcccagagtt
     2461 ccactttttc ctgaaaatgg tttacaccag cagccagaac ccttgcttcc aaataatatg
     2521 aaatctgcct gtgaaaaacg tttaggatgt tgtagttctc ctcattctaa gccaaattgc
     2581 tcaacccttt ctccaccaat gccactgccc cagctgttac cttcggttac tgatgcaagg
     2641 tcggcaggac cttctgatca tattgattcc tcagttactg gggttcaaag gtttcgagat
     2701 actctaaaaa taccctacaa gctggaatta aaaaatgaac cagggagaac ggatttgaaa
     2761 cacattgtta tagatgggag caatgttgca attacccatg gtctgaaaaa gttcttttct
     2821 tgtcgtggaa ttgcaattgc agttgaatat ttttggaagc ttggcaacag aaacatcact
     2881 gtatttgtcc ctcagtggag aacaaggcgt gatcctaatg tcacagaaca gcacttctta
     2941 acccagctcc aggagctcgg aatattatct ttaactcctg cccggatggt ctttggagaa
     3001 agaattgctt ctcatgatga caggtttcta ctacacttag cggacaaaac tggtggcata
     3061 attgtgacaa atgataattt cagagaattt gtgaatgagt cagtctcttg gagagaaatt
     3121 attacaaaaa ggctgctgca gtacacgttc gtgggggaca tatttatggt tcccgatgat
     3181 cctctgggaa gaagtggacc tcgattagag gagtttcttc agaaggaagt ctgtcttaga
     3241 gatatgcagc ccctactcag tgccctgcca aatgtgggca tgtttgaccc cagcttcaga
     3301 gtccctggca cccaggcagc cagcaccagc caccagcctc cgacccggat tcagggagcc
     3361 ccttcaagcc actggctccc tcagcagccc cacttcccac tgctgccagc cctccccagt
     3421 ctccagcaga acctgcccat gccagctcag agatcttctg cagaaaccaa cgagctgagg
     3481 gaagcccttc tgaagatctt ccctgactca gagcaaagac tgaaaataga ccagatcttg
     3541 gtggcccacc catacatgaa agatctaaat gcgctttctg ccatggtgtt ggatctcgag
     3601 tctagagact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     3661 gacgatgaca agggaagcgg agccactaac ttctccctgt tgaaacaagc aggggatgtc
     3721 gaagagaatc ccgggccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     3781 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3841 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3901 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3961 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4021 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4081 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4141 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4201 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     4261 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4321 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     4381 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4441 actctcggca tggacgagct gtacaagtaa gggcccgttt aaacccgctg atcagcctcg
     4501 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4561 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4621 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4681 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4741 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4801 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4861 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4921 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4981 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5041 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5101 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5161 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5221 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5281 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5341 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5401 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5461 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5521 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5581 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5641 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5701 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5761 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5821 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5881 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5941 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6001 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6061 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6121 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6181 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6241 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6301 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6361 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6421 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6481 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6541 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6721 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7561 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7681 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7741 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7801 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7861 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7921 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7981 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8041 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8101 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8161 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8221 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8281 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8341 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8401 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8461 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8521 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8581 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8641 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8701 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8761 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8821 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8881 ccgaaaagtg ccacctgacg tc
//
