LOCUS       V039971                 7197 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039971
VERSION     V039971
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1877
                     /label="TMEFF2(NM_016192)"
                     /note="TMEFF2(NM_016192)"
                     /gene="TMEFF2"
     CDS             1884..1949
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1973..2561
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2627..2675
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2877..3305
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3319..3648
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3715..4509
                     /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    4685..4806
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4855..4871)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4879..4895
                     /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(4903..4933)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4948..4969
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5257..5845)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6016..6876)
                     /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(6877..6981)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt gctgtgggag tccccgcggc
      781 agtgcagcag ctggacactt tgcgagggct tttgctggct gctgctgctg cccgtcatgc
      841 tactcatcgt agcccgcccg gtgaagctcg ctgctttccc tacctcctta agtgactgcc
      901 aaacgcccac cggctggaat tgctctggtt atgatgacag agaaaatgat ctcttcctct
      961 gtgacaccaa cacctgtaaa tttgatgggg aatgtttaag aattggagac actgtgactt
     1021 gcgtctgtca gttcaagtgc aacaatgact atgtgcctgt gtgtggctcc aatggggaga
     1081 gctaccagaa tgagtgttac ctgcgacagg ctgcatgcaa acagcagagt gagatacttg
     1141 tggtgtcaga aggatcatgt gccacagatg caggatcagg atctggagat ggagtccatg
     1201 aaggctctgg agaaactagt caaaaggaga catccacctg tgatatttgc cagtttggtg
     1261 cagaatgtga cgaagatgcc gaggatgtct ggtgtgtgtg taatattgac tgttctcaaa
     1321 ccaacttcaa tcccctctgc gcttctgatg ggaaatctta tgataatgca tgccaaatca
     1381 aagaagcatc gtgtcagaaa caggagaaaa ttgaagtcat gtctttgggt cgatgtcaag
     1441 ataacacaac tacaactact aagtctgaag atgggcatta tgcaagaaca gattatgcag
     1501 agaatgctaa caaattagaa gaaagtgcca gagaacacca cataccttgt ccggaacatt
     1561 acaatggctt ctgcatgcat gggaagtgtg agcattctat caatatgcag gagccatctt
     1621 gcaggtgtga tgctggttat actggacaac actgtgaaaa aaaggactac agtgttctat
     1681 acgttgttcc cggtcctgta cgatttcagt atgtcttaat cgcagctgtg attggaacaa
     1741 ttcagattgc tgtcatctgt gtggtggtcc tctgcatcac aaggaaatgc cccagaagca
     1801 acagaattca cagacagaag caaaatacag ggcactacag ttcagacaat acaacaagag
     1861 cgtccacgag gttaatcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1921 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1981 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2041 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2101 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2161 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2221 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2281 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2341 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2401 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2461 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2521 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2581 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2641 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2701 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2761 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2821 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2881 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2941 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3001 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3061 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3121 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3181 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3241 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3301 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3361 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3421 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3481 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3541 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3601 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3661 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3721 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3781 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3841 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3901 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3961 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4021 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4081 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4141 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4201 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4261 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4321 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4381 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4441 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4501 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4561 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4621 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4681 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4741 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4801 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4861 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4921 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5761 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7021 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     7081 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7141 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
