LOCUS       V018358                 7926 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018358
VERSION     V018358
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             922..939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             940..3414
                     /label="HNRNPU(NM_031844)"
                     /note="HNRNPU(NM_031844)"
                     /gene="HNRNPU"
     promoter        complement(3478..3496)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    3522..3746
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3792..4220
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4234..4564
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4631..5425
                     /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    5599..5720
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5769..5785)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5793..5809
                     /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(5817..5847)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5862..5883
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6171..6759)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6930..7790)
                     /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(7791..7895)
                     /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 gcttggtacc
      901 gagctcggat ccgccaccat gcatcaccat caccatcaca tgagttcctc gcctgttaat
      961 gtaaaaaagc tgaaggtgtc ggagctgaaa gaggagctca agaagcgacg cctttctgac
     1021 aagggtctca aggccgagct catggagcga ctccaggctg cgctggacga cgaggaggcc
     1081 gggggccgcc ccgccatgga gcccgggaac ggcagcctag acctgggcgg ggattccgct
     1141 gggcgctcgg gagcaggcct cgagcaggag gccgcggccg gcggcgatga agaggaggag
     1201 gaagaggaag aggaggagga aggaatctcc gctctggacg gcgaccagat ggagctagga
     1261 gaggagaacg gggccgcggg ggcggccgac tcgggcccga tggaggagga ggaggccgcc
     1321 tcggaagacg agaacggcga cgatcagggt ttccaggaag gggaagatga gctcggggac
     1381 gaagaggaag gcgcgggcga cgagaacggg cacggggagc agcagcctca accgccggcg
     1441 acgcagcagc aacagcccca acagcagcgc ggggccgcca aggaggccgc ggggaagagc
     1501 agcggcccca cctcgctgtt cgcggtgacg gtggcgccgc ccggggcgag gcagggccag
     1561 cagcaggcgg gaggtaagaa gaaggcggaa ggcggcggag gcggcggtcg ccccggggct
     1621 ccggcggcgg gggacggcaa aacagaacag aaaggcggag ataaaaagag gggtgttaaa
     1681 agaccacgag aagatcatgg ccgtggatat tttgagtaca ttgaagagaa caagtatagc
     1741 agagccaaat ctcctcagcc acctgttgaa gaagaagatg aacacttcga tgacacagtg
     1801 gtttgtcttg atacttataa ttgtgatcta cattttaaaa tatcaagaga tcgtctcagt
     1861 gcttcttccc ttacaatgga gagttttgct tttctttggg ctggaggaag agcatcctat
     1921 ggtgtgtcaa aaggcaaagt gtgttttgag atgaaggtta cagagaagat cccagtaagg
     1981 catttatata caaaagatat tgacatacat gaagttcgta ttggctggtc actaactaca
     2041 agtggaatgt tacttggtga agaagaattt tcttatgggt attctctaaa aggaataaaa
     2101 acatgcaact gtgagactga agattatgga gaaaagtttg atgaaaatga tgtgattaca
     2161 tgttttgcta actttgaaag tgatgaagta gaactctcgt atgctaagaa tggacaagat
     2221 cttggcgttg ccttcaaaat cagtaaggaa gttcttgctg gacggccact gttcccgcat
     2281 gttctctgcc acaactgtgc agttgaattt aattttggtc agaaggaaaa gccatatttt
     2341 ccaatacctg aagagtatac tttcatccag aacgtcccct tagaggatcg agttagagga
     2401 ccaaaggggc ctgaagagaa gaaagattgt gaagttgtga tgatgattgg cttgccagga
     2461 gctggaaaaa ctacctgggt tactaaacat gcagcagaaa atccagggaa atataacatt
     2521 cttggcacaa atactattat ggataagatg atggtggcag gttttaagaa gcaaatggca
     2581 gatactggaa aactgaacac actgttgcag agagcccccc agtgtcttgg gaaatttatt
     2641 gagattgctg cccgaaagaa gcgaaatttt attctggatc agacaaatgt gtctgctgct
     2701 gcccagagga gaaaaatgtg cctgtttgca ggcttccagc gaaaagctgt tgtagtttgc
     2761 ccaaaagatg aagactataa gcaaagaaca cagaagaaag cagaagtaga ggggaaagac
     2821 ctaccagaac atgcggtcct caaaatgaaa ggaaacttta ccctcccaga ggtagctgag
     2881 tgctttgatg aaataaccta tgttgaactt cagaaggaag aagcccaaaa actcttggag
     2941 caatataagg aagaaagcaa aaaggctctt ccaccagaaa agaaacagaa cactggctca
     3001 aagaaaagca ataaaaataa gagtggcaag aaccagttta acagaggtgg tggccataga
     3061 ggacgtggag gattcaatat gcgtggtgga aatttcagag gaggagcccc tgggaatcgt
     3121 ggcggatata ataggagggg caacatgcca cagagaggtg gtggcggtgg aggaagtggt
     3181 ggaatcggct atccataccc tcgtgcccct gtttttcctg gccgtggtag ttactcaaac
     3241 agagggaact acaacagagg tggaatgccc aacagaggga actacaacca gaacttcaga
     3301 ggacgaggaa acaatcgtgg ctacaaaaat caatctcagg gctacaacca gtggcagcag
     3361 ggtcaattct ggggtcagaa gccatggagt cagcattatc accaaggata ttattgagaa
     3421 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg gccctattct
     3481 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     3541 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3601 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3661 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3721 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3781 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3841 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3901 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3961 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4021 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4081 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4141 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4201 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4261 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4501 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4561 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4621 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4681 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4741 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4801 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4861 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4921 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4981 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5041 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5101 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5161 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5221 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5281 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5341 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5401 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5461 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5521 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5581 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5641 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5701 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6361 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6601 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7921 gacgtc
//
