LOCUS       V031797                 8107 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031797
VERSION     V031797
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             973..999
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1000..1023
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1024..3627
                     /label="Gpr155(NM_001190297)"
                     /note="Gpr155(NM_001190297)"
                     /gene="Gpr155"
     polyA_signal    3674..3898
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3944..4372
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4386..4715
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4782..5576
                     /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    5750..5871
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5920..5936)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5944..5960
                     /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(5968..5998)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6013..6034
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6322..6907)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7078..7938)
                     /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(7939..8043)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcatgaag accatcatcg ccctgagcta catcttctgc
      961 ctggtgttcg cctacccata cgacgtccca gactacgctg actacaagga cgatgatgac
     1021 aaaatggatt cttatttctc agcaaagaac tcaacgcttg ctggtgatat gaatgcgact
     1081 tggcctgcct ctcatggatt taatgccacc ggcgatccac cttcgatgtc gataacccgg
     1141 ctcttcccgg ccttactgga gtgctttggc atagtccttt gtggctacat agcaggaagg
     1201 gccaatatca taacctcaac acaggccaaa ggactgggga atttcgtctc cagatttgcc
     1261 cttccagcct tattattcaa aaacatggtt gtgcttaatt tttccaacgt ggactgggct
     1321 tttctctaca gcgtcttaat tggcaaagcc tccgtgtttt tcattgtgtg tgtattaacc
     1381 ttattggtgg ccagtcctga gagccggttt agcaaagctg gactattccc tatttttgct
     1441 acacagagta atgactttgc tttgggatac cctatagttg aagctctgta ccaatctaca
     1501 tatccagagt atctgcagta catttatttg gtggcaccaa tatctcttat gatgctaaac
     1561 cccatagggt ttatcttctg tgaaatccag aagtcaaaag acactcaaaa tgcttctcaa
     1621 aataaagcaa aaattgtggg acttggattt ctacgcgtgt tacagaaccc gatagtattt
     1681 atggtctttg ttggcattgc cttcaatttt attcttgata aaaagatccc agtatatatg
     1741 gaaaactttc tggatgggct tgcaaattcc ttctctggat cagccctgtt ttatcttggt
     1801 ctgaccatgg taggaaaaat caggcgactg aagaagtcgg cgtttgtggt tcttactctt
     1861 ctcatcacag ccaaactcct ggtgctgccc ctcctgtgca gagaaatggt ggaactcctg
     1921 gacaaggggg acagcgtagt gaaccataca agcctgtcca actatgcgtt tctatatggt
     1981 gtcttccctg tggctccagg agtggctatc tttgcaacac agttcaacat ggaagtggag
     2041 attataacct caggcatggt aataagtaca tttgtgtctg ctccaatcat gtatgtttct
     2101 gcctggttat taacctttcc taccatggat gccaagccac tggcgtatgc cattcagaat
     2161 gttagctttg atataagtat tatcagcctg gtctccttga tctggtccct gtctattctt
     2221 ctcttgagta agaaatataa gcagctccca catatgctca cagcgaactt actcatcgct
     2281 cagactattg tttgtgctgg aatgatgata tggaattttg ttaaagaaaa aaattttgtt
     2341 ggacaaattt tggtgtttgt tttattgtac agctccctgt acagcaccta tctgtggaca
     2401 ggccttctag cagtttcttt gtttctttta aagaagcgcg agagcgttca gctcccggtg
     2461 ggaatcatca tcatctctgg ctgggggatc cctgctctcc ttgtgggcgt gcttctgata
     2521 actggaaaac acaacggaga cagcattgac tcagccttct tttatggaaa agagcagatg
     2581 atcaccacgg cagtcaccct gttctgcagc attctcatag ctggggtgtc actcatgtgc
     2641 atgaaccgca ccacccaagc tggtcactac gaaggtttcg gccagtctca gaaccacaaa
     2701 ccagtagagc ctggaagcac tgcgtttgag gagaacccag caccaacaaa tgaaccagaa
     2761 ctcttcccaa gttctatccc agaaacaagt tgctgttctt gctccttggg aaatggtgaa
     2821 ttacgctgcc catccataga gccagtaacg aactcaagtg ctagtgggcc tatgccttct
     2881 tcctttgaga aaaccgacca ctgtgtgagc cgttgtgact cccagagctg catcctcgcc
     2941 caggaagaag agcagtacct gcagagcgga gacccacagc tgacccggca cgtcctgttg
     3001 tgcttgctgc ttatcatcgg cctctttgct aatctctcca gctgcctgtg gtggttgttc
     3061 aaccacgaga cgggacggct ctatgttgaa ttacagtttt tctgtgccgt gtttaacttt
     3121 ggccagggat ttatttcctt tggaatcttt gggctggaca aacatctaat catcctacct
     3181 ttcaaaagaa ggcttgaatt cctgtggaac aacaaagaag cagcagcaga cagagaatcc
     3241 cccgtttccg aggaaataaa aatgacctgt caacagtttg tgcattatca ccgcgacctg
     3301 tgcatccgaa acatcgtcag agaaaggagg tgtggtgcca agacttctgc cgggaccttc
     3361 tgtggctgtg acctggtgaa ctggctcatc gaagtcggcc ttgcttctga ccgcggcgaa
     3421 gctgtgatct acggagacag actggtgcaa gggggggtca tccaacacat caccaatgaa
     3481 tacgagttcc gggacgagta tctcttttac cgattccttc agaagagtcc tgagcagagt
     3541 cctcctgccc gtactctaag agaccaccag gaggaaagct ataaagaaat cggtcactca
     3601 tctccaccct cagtctcccc taagacttaa ctcgagtcta gagggcccgt ttaaacccgc
     3661 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3721 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3781 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3841 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3901 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3961 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4021 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4081 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4141 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4201 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4261 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4321 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4381 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4441 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4561 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4621 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4681 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4741 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4801 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4861 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4921 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4981 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5041 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5101 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5161 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5221 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5281 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5341 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5401 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5461 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5521 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5581 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5641 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5701 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5881 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5941 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6001 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6121 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6841 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8101 ctatggt
//
