LOCUS       V045109                 7282 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045109
VERSION     V045109
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             706..1986
                     /label="FLOT1(NM_005803)"
                     /note="FLOT1(NM_005803)"
                     /gene="FLOT1"
     CDS             1993..2034
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     misc_feature    2058..2646
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2712..2760
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2962..3390
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3404..3733
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3800..4594
                     /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    4770..4891
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4940..4956)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4964..4980
                     /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(4988..5018)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5033..5054
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5342..5930)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6101..6961)
                     /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(6962..7066)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgtt tttcacttgt
      721 ggcccaaatg aggccatggt ggtctccggg ttctgccgaa gccccccagt catggtggct
      781 ggagggcgtg tctttgtcct gccctgcatc caacagatcc agaggatctc tctcaacaca
      841 ctgaccctca atgtcaagag tgaaaaggtt tacactcgcc atggggtccc catctcagtc
      901 actggcattg cccaggtaaa aatccagggg cagaacaagg agatgttggc ggccgcctgt
      961 cagatgttcc tggggaagac ggaggctgag attgcccaca ttgccctgga gacgttagag
     1021 ggccaccaga gggccatcat ggcccacatg actgtggagg agatctataa ggacaggcag
     1081 aaattctcag aacaggtttt caaagtggcc tcctcagacc tggtcaacat gggcatcagt
     1141 gtggttagct acactctgaa ggacattcac gatgaccagg actatttgca ctctttgggg
     1201 aaggctcgaa cagctcaagt ccaaaaagat gcacggattg gagaagcaga ggccaagaga
     1261 gatgctggga tccgggaagc taaagccaag caggaaaagg tgtctgctca gtacctgagt
     1321 gagatcgaga tggccaaggc acagagagat tacgaactga agaaggccgc ctatgacatc
     1381 gaggtcaaca cccgccgagc acaggctgac ctggcctatc agcttcaggt ggccaagact
     1441 aagcagcaga ttgaggagca gcgggtgcag gtgcaggtgg tggagcgggc ccagcaggtg
     1501 gcagtgcagg agcaggagat cgcccggcgg gagaaggagc tggaggcccg ggtgcggaag
     1561 ccagcggaag cggagcgcta caagctggag cgcctagccg aggcagagaa gtcccaacta
     1621 attatgcagg cggaggcaga agccgcgtct gtgcggatgc gtggggaagc tgaggccttt
     1681 gccatagggg cccgagcccg agccgaggct gagcagatgg ccaagaaggc agaagccttc
     1741 cagctgtacc aagaggctgc tcagctggac atgctgctag agaagctgcc ccaggtggca
     1801 gaggagatca gtggtccctt gacttcagcc aataagatca cactggtgtc cagcggcagt
     1861 gggaccatgg gggcagccaa agtgactggg gaagtactgg acattctaac tcgcctgcca
     1921 gagagtgtgg aaagactcac aggcgtgagc atctcccagg tgaatcacaa gcctttgaga
     1981 acagccctcg agggtaagcc tatccctaac cctctcctcg gtctcgattc tacgtagtga
     2041 gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2101 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2161 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2221 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2281 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2341 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2401 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     2461 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     2521 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2581 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2641 ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     2701 ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc
     2761 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     2821 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     2881 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct
     2941 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3001 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3061 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     3121 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3181 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3241 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3301 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     3361 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3481 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3541 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3601 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3661 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3721 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3781 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3841 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3901 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3961 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4021 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4081 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4141 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4201 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4261 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4321 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4381 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4441 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4501 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4561 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     4621 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4681 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4741 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4861 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4921 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4981 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5041 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5101 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5161 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5221 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5281 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5341 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5401 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5461 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5521 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5581 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5641 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5701 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5761 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5821 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5941 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6001 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6061 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6121 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6181 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6241 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6301 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6361 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6421 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6481 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6541 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6601 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6661 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6721 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6781 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6841 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6901 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7021 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7081 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag
     7141 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7201 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     7261 ttgcatgaag aatctgctta gg
//
