LOCUS       V020645                 6782 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020645
VERSION     V020645
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..1462
                     /label="UPK1B(NM_006952)"
                     /note="UPK1B(NM_006952)"
                     /gene="UPK1B"
     CDS             1469..1534
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1558..2146
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2212..2260
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2462..2890
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2904..3233
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3300..4094
                     /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    4270..4391
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4440..4456)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4464..4480
                     /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(4488..4518)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4533..4554
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4842..5430)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5601..6461)
                     /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(6462..6566)
                     /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 ccatggccaa agacaactca actgttcgtt gcttccaggg
      721 cctgctgatt tttggaaatg tgattattgg ttgttgcggc attgccctga ctgcggagtg
      781 catcttcttt gtatctgacc aacacagcct ctacccactg cttgaagcca ccgacaacga
      841 tgacatctat ggggctgcct ggatcggcat atttgtgggc atctgcctct tctgcctgtc
      901 tgttctaggc attgtaggca tcatgaagtc cagcaggaaa attcttctgg cgtatttcat
      961 tctgatgttt atagtatatg cctttgaagt ggcatcttgt atcacagcag caacacaaca
     1021 agactttttc acacccaacc tcttcctgaa gcagatgcta gagaggtacc aaaacaacag
     1081 ccctccaaac aatgatgacc agtggaaaaa caatggagtc accaaaacct gggacaggct
     1141 catgctccag gacaattgct gtggcgtaaa tggtccatca gactggcaaa aatacacatc
     1201 tgccttccgg actgagaata atgatgctga ctatccctgg cctcgtcaat gctgtgttat
     1261 gaacaatctt aaagaacctc tcaacctgga ggcttgtaaa ctaggcgtgc ctggttttta
     1321 tcacaatcag ggctgctatg aactgatctc tggtccaatg aaccgacacg cctggggggt
     1381 tgcctggttt ggatttgcca ttctctgctg gactttttgg gttctcctgg gtaccatgtt
     1441 ctactggagc agaattgaat atctcgagga ctacaaagac catgacggtg attataaaga
     1501 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     1561 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1621 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1681 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     1741 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     1801 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     1861 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1921 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     1981 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     2041 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     2101 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     2161 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     2221 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     2281 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     2341 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     2401 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     2461 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2581 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2821 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2881 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3241 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3301 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3361 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3421 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3481 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3541 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3601 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3661 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3721 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3781 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3841 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3901 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3961 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4021 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4081 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     4141 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4201 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4261 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4381 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4441 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4501 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4561 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4621 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4681 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4741 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4801 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4861 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4921 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4981 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     5041 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5101 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5161 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5221 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5281 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5341 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5401 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5461 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5521 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5581 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5641 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5701 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5761 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5821 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5881 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5941 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6001 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6061 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6121 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6181 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6241 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6301 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6361 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6421 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6481 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6541 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6601 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     6661 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     6721 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6781 gg
//
