LOCUS       V040538                 7330 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040538
VERSION     V040538
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             928..2766
                     /label="GPR37(NM_005302)"
                     /note="GPR37(NM_005302)"
                     /gene="GPR37"
     CDS             2773..2796
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2803..2892
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    2930..3154
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3200..3628
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3642..3971
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4038..4832
                     /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    5006..5127
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5176..5192)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5200..5216
                     /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(5224..5254)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5269..5290
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5578..6163)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6334..7194)
                     /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(7195..7299)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctgaatt cgccaccatg cgagccccgg gcgcgcttct cgcccgcatg
      961 tcgcggctac tgcttctgct actgctcaag gtgtctgcct cttctgccct cggggtcgcc
     1021 cctgcgtcca gaaacgaaac ttgtctgggg gagagctgtg cacctacagt gatccagcgc
     1081 cgcggcaggg acgcctgggg accgggaaat tctgcaagag acgttctgcg agcccgagca
     1141 cccagggagg agcagggggc agcgtttctt gcgggaccct cctgggacct gccggcggcc
     1201 ccgggccgtg acccggctgc aggcagaggg gcggaggcgt cggcagccgg acccccggga
     1261 cctccaacca ggccacctgg cccctggagg tggaaaggtg ctcggggtca ggagccttct
     1321 gaaactttgg ggagagggaa ccccacggcc ctccagctct tccttcagat ctcagaggag
     1381 gaagagaagg gtcccagagg cgctggcatt tccgggcgta gccaggagca gagtgtgaag
     1441 acagtccccg gagccagcga tcttttttac tggccaagga gagccgggaa actccagggt
     1501 tcccaccaca agcccctgtc caagacggcc aatggactgg cggggcacga agggtggaca
     1561 attgcactcc cgggccgggc gctggcccag aatggatcct tgggtgaagg aatccatgag
     1621 cctgggggtc cccgccgggg aaacagcacg aaccggcgtg tgagactgaa gaaccccttc
     1681 tacccgctga cccaggagtc ctatggagcc tacgcggtca tgtgtctgtc cgtggtgatc
     1741 ttcgggaccg gcatcattgg caacctggcg gtgatgtgca tcgtgtgcca caactactac
     1801 atgcggagca tctccaactc cctcttggcc aacctggcct tctgggactt tctcatcatc
     1861 ttcttctgcc ttccgctggt catcttccac gagctgacca agaagtggct gctggaggac
     1921 ttctcctgca agatcgtgcc ctatatagag gtcgcttctc tgggagtcac cactttcacc
     1981 ttatgtgctc tgtgcataga ccgcttccgt gctgccacca acgtacagat gtactacgaa
     2041 atgatcgaaa actgttcctc aacaactgcc aaacttgctg ttatatgggt gggagctcta
     2101 ttgttagcac ttccagaagt tgttctccgc cagctgagca aggaggattt ggggtttagt
     2161 ggccgagctc cggcagaaag gtgcattatt aagatctctc ctgatttacc agacaccatc
     2221 tatgttctag ccctcaccta cgacagtgcg agactgtggt ggtattttgg ctgttacttt
     2281 tgtttgccca cgcttttcac catcacctgc tctctagtga ctgcgaggaa aatccgcaaa
     2341 gcagagaaag cctgtacccg agggaataaa cggcagattc aactagagag tcagatgaac
     2401 tgtacagtag tggcactgac cattttatat ggattttgca ttattcctga aaatatctgc
     2461 aacattgtta ctgcctacat ggctacaggg gtttcacagc agacaatgga cctccttaat
     2521 atcatcagcc agttcctttt gttctttaag tcctgtgtca ccccagtcct ccttttctgt
     2581 ctctgcaaac ccttcagtcg ggccttcatg gagtgctgct gctgttgctg tgaggaatgc
     2641 attcagaagt cttcaacggt gaccagtgat gacaatgaca acgagtacac cacggaactc
     2701 gaactctcgc ctttcagtac catacgccgt gaaatgtcca cttttgcttc tgtcggaact
     2761 cattgcctcg aggattacaa ggatgacgac gataagggat ccagcgcctg gtcccaccct
     2821 cagtttgaga agggcggagg ctctggcggc ggaagcggag gatctgcttg gagccacccc
     2881 cagttcgaaa agtgatgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     2941 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3001 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3061 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3121 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3181 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3241 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3301 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3361 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3421 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3481 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3541 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3601 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3661 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3721 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3781 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3841 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3901 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3961 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4021 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4081 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4141 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4201 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4261 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4321 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4381 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4441 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4501 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4561 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4621 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4681 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4741 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4801 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4861 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4921 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4981 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5101 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5161 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5221 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5281 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5341 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5401 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5461 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5521 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5581 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5641 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5701 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5761 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5821 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5881 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5941 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6001 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6061 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6541 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6601 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6661 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6721 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6781 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6841 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6901 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6961 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7021 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7081 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7141 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7201 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7261 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7321 acctgacgtc
//
