LOCUS       V039059                 7942 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039059
VERSION     V039059
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..2598
                     /label="RAD21(NM_006265)"
                     /note="RAD21(NM_006265)"
                     /gene="RAD21"
     CDS             2605..2694
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2718..3306
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3372..3420
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3622..4050
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4064..4393
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4460..5254
                     /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    5430..5551
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5600..5616)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5624..5640
                     /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(5648..5678)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5693..5714
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6002..6590)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6761..7621)
                     /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(7622..7726)
                     /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 ctacgcacat
      721 tttgttctca gtaaaagagg gcctctggcc aaaatttggc tagcggccca ttgggataag
      781 aagctaacca aagcccatgt gttcgagtgt aatttagaga gcagcgtgga gagtatcatc
      841 tcaccaaagg tgaaaatggc attacggaca tcaggacatc tcttactggg agtagttcga
      901 atctatcaca ggaaagccaa ataccttctt gcagactgta atgaagcatt cattaagata
      961 aagatggctt ttcggccagg tgtggttgac ctgcctgagg aaaatcggga agcagcttat
     1021 aatgccatta ctttacctga agaatttcat gactttgatc agccactgcc tgacttagat
     1081 gacatcgatg tggcccagca gttcagcttg aatcagagta gagtggaaga gataaccatg
     1141 agagaagaag ttgggaacat cagtatttta caagaaaatg attttggtga ttttggaatg
     1201 gatgatcgtg agataatgag agaaggcagt gcttttgagg atgacgacat gttagtaagc
     1261 actactactt ctaacctcct attagagtct gaacagagca ccagcaatct gaatgagaaa
     1321 attaaccatt tagaatatga agatcaatat aaggatgata attttggaga aggaaatgat
     1381 ggtggaatat tagatgacaa acttattagt aataatgatg gcggtatctt tgatgatccc
     1441 cctgccctct ctgaggcagg ggtgatgttg ccagagcagc ctgcacatga cgatatggat
     1501 gaggatgata atgtatcaat gggtgggcct gatagtcctg attcagtgga tcccgttgaa
     1561 ccaatgccaa ccatgactga tcaaacaaca cttgttccaa atgaggaaga agcatttgca
     1621 ttggagccta ttgatataac tgttaaagaa acaaaagcca agaggaagag gaagctaatt
     1681 gttgacagtg tcaaagagtt ggatagcaag acaattagag cccaacttag tgattattca
     1741 gatattgtta ctactttgga tctggcaccg cccaccaaga aattgatgat gtggaaagag
     1801 acaggaggag tagaaaaact gttttcttta cctgctcagc ctttgtggaa taacagacta
     1861 ctgaagctct ttacacgctg tcttacaccg cttgtaccag aagaccttag aaaaaggagg
     1921 aaaggaggag aggcagataa tttggatgaa ttcctcaaag aatttgaaaa tccagaggtt
     1981 cctagagagg accagcaaca gcagcatcag cagcgtgatg ttatcgatga gcccattatt
     2041 gaagagccaa gccgcctcca ggagtcagtg atggaggcca gcagaacaaa catagatgag
     2101 tcagctatgc ctccaccacc acctcaggga gttaagcgaa aagctggaca aattgaccca
     2161 gagcctgtga tgcctcctca gcaggtagag cagatggaaa taccacctgt agagcttccc
     2221 ccagaagaac ctccaaatat ctgtcagcta ataccagagt tagaacttct gccagaaaaa
     2281 gagaaggaga aagagaagga aaaagaagat gatgaagagg aagaggatga agatgcatca
     2341 gggggcgatc aagatcagga agaaagaaga tggaacaaaa ggactcagca gatgcttcat
     2401 ggtcttcagc gtgctcttgc taaaactgga gctgaatcta tcagtttgct tgagttatgt
     2461 cgaaatacga acagaaaaca agctgccgca aagttctaca gcttcttggt tcttaaaaag
     2521 cagcaagcta ttgagctgac acaggaagaa ccgtacagtg acatcatcgc aacacctgga
     2581 ccaaggttcc atattatact cgaggaacaa aaactaatat cagaggaaga tttggaacag
     2641 aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttatagtga
     2701 gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2761 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2821 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2881 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2941 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3001 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3061 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     3121 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3181 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3241 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3301 ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3361 ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc
     3421 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3481 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3541 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct
     3601 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3661 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3721 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     3781 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3841 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3901 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3961 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     4021 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4081 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4141 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4321 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4381 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4441 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4501 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4561 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4621 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4681 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4741 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4801 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4861 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4921 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4981 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5041 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5101 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5161 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5221 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     5281 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5341 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5401 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5521 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5581 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5641 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5701 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5761 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6181 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6421 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7741 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag
     7801 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7861 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     7921 ttgcatgaag aatctgctta gg
//
