LOCUS       V035013                 7267 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035013
VERSION     V035013
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..1926
                     /label="TMEM184B(NM_012264)"
                     /note="TMEM184B(NM_012264)"
                     /gene="TMEM184B"
     CDS             1933..1959
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1963..1989
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1993..2019
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2043..2631
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2697..2745
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2947..3375
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3389..3718
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3785..4579
                     /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    4755..4876
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4925..4941)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4949..4965
                     /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(4973..5003)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5018..5039
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5327..5915)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6086..6946)
                     /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(6947..7051)
                     /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 ccaccatgac agtgaggggg
      721 gatgtgctgg ccccggatcc agcgtcgccc acgaccgcag cagcctcgcc cagcgtctcc
      781 gtgatccccg agggcagccc cactgccatg gagcagcctg tgttcctgat gacaactgcc
      841 gctcaggcca tctctggctt cttcgtgtgg acggccctgc tcatcacatg ccaccagatc
      901 tacatgcacc tgcgctgcta cagctgcccc aacgagcagc gctacatcgt gcgcatcctc
      961 ttcatcgtgc ccatctacgc ctttgactcc tggctcagcc tcctcttctt caccaacgac
     1021 cagtactacg tgtacttcgg caccgtccgc gactgctatg aggccttggt catctataat
     1081 ttcctgagcc tgtgctatga gtacctagga ggagaaagtt ccatcatgtc ggagatcaga
     1141 ggaaaaccca ttgagtccag ctgtatgtat ggcacctgct gcctctgggg aaagacttat
     1201 tccatcggat ttctgaggtt ctgcaaacag gccaccctgc agttctgtgt ggtgaagcca
     1261 ctcatggcgg tcagcactgt ggtcctccag gccttcggca agtaccggga tggggacttt
     1321 gacgtcacca gtggctacct ctacgtgacc atcatctaca acatctccgt cagcctggcc
     1381 ctctacgccc tcttcctctt ctacttcgcc acccgggagc tgctcagccc ctacagcccc
     1441 gtcctcaagt tcttcatggt caagtccgtc atctttcttt ccttctggca aggcatgctc
     1501 ctggccatcc tggagaagtg tggggccatc cccaaaatcc actcggcccg cgtgtcggtg
     1561 ggcgagggca ccgtggctgc cggctaccag gacttcatca tctgtgtgga gatgttcttt
     1621 gcagccctgg ccctgcggca cgccttcacc tacaaggtct atgctgacaa gaggctggac
     1681 gcacaaggcc gctgtgcccc catgaagagc atctccagca gcctcaagga gaccatgaac
     1741 ccgcacgaca tcgtgcagga cgccatccac aacttctcac ctgcctacca gcagtacacg
     1801 cagcagtcca ccctggagcc tgggcccacc tggcgtggtg gcgcccacgg cctctcccgc
     1861 tcccacagcc tcagtggcgc ccgcgacaac gagaagactc tcctgctcag ctctgatgat
     1921 gaattcctcg agtacccata cgacgtacct gattatgcaa cttatcctta tgatgtacct
     1981 gattatgcta gctatcctta tgatgtccca gactacgcat agtgagggcc cgatatctcg
     2041 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2101 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2161 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2221 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2281 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2341 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2401 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     2461 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2521 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2581 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg
     2641 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     2701 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc
     2761 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     2821 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     2881 ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt
     2941 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3001 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3061 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc
     3121 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3181 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3241 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3301 atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3361 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3421 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3541 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3601 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3661 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3721 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3781 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3841 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3901 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3961 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4021 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4081 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4141 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4201 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4261 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4321 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4381 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4441 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4501 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4561 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     4621 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4681 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4741 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4801 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4861 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4921 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4981 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5041 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5101 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5161 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5221 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5281 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5341 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5401 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5461 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5521 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5581 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5641 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5701 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5761 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5821 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5881 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5941 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6001 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6061 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6121 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6181 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6241 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6301 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6361 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6421 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6481 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6541 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6601 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6661 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6721 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6781 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6841 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6901 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6961 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7021 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7081 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
     7141 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     7201 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     7261 gcttagg
//
