LOCUS       V040035                 6691 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040035
VERSION     V040035
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..2127
                     /label="DRD3(NM_000796)"
                     /note="DRD3(NM_000796)"
                     /gene="DRD3"
     CDS             2134..2157
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2164..2253
                     /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    2291..2515
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2561..2989
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3003..3332
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3399..4193
                     /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    4367..4488
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4537..4553)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4561..4577
                     /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(4585..4615)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4630..4651
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4939..5524)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5695..6555)
                     /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(6556..6660)
                     /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 gcatctctga gccagctgag tggccacctg
      961 aactacacct gtggggcaga gaactccaca ggtgccagcc aggcccgccc acatgcctac
     1021 tatgccctct cctactgcgc gctcatcctg gccatcgtct tcggcaatgg cctggtgtgc
     1081 atggctgtgc tgaaggagcg ggccctgcag actaccacca actacttagt agtgagcctg
     1141 gctgtggcag acttgctggt ggccaccttg gtgatgccct gggtggtata cctggaggtg
     1201 acaggtggag tctggaattt cagccgcatt tgctgtgatg tttttgtcac cctggatgtc
     1261 atgatgtgta cagccagcat ccttaatctc tgtgccatca gcatagacag gtacactgca
     1321 gtggtcatgc ccgttcacta ccagcatggc acgggacaga gctcctgtcg gcgcgtggcc
     1381 ctcatgatca cggccgtctg ggtactggcc tttgctgtgt cctgccctct tctgtttggc
     1441 tttaatacca caggggaccc cactgtctgc tccatctcca accctgattt tgtcatctac
     1501 tcttcagtgg tgtccttcta cctgcccttt ggagtgactg tccttgtcta tgccagaatc
     1561 tatgtggtgc tgaaacaaag gagacggaaa aggatcctca ctcgacagaa cagtcagtgc
     1621 aacagtgtca ggcctggctt cccccagcaa accctctctc ctgacccggc acatctggag
     1681 ctgaagcgtt actacagcat ctgccaggac actgccttgg gtggaccagg cttccaagaa
     1741 agaggaggag agttgaaaag agaggagaag actcggaatt ccctgagtcc caccatagcg
     1801 cccaagctca gcttagaagt tcgaaaactc agcaatggca gattatcgac atctttgaag
     1861 ctggggcccc tgcaacctcg gggagtgcca cttcgggaga agaaggcaac ccaaatggtg
     1921 gccattgtgc ttggggcctt cattgtctgc tggctgccct tcttcttgac ccatgttctc
     1981 aatacccact gccagacatg ccacgtgtcc ccagagcttt acagtgccac gacatggctg
     2041 ggctacgtga atagcgccct caaccctgtg atctatacca ccttcaatat cgagttccgg
     2101 aaagccttcc tcaagatcct gtcttgcctc gaggattaca aggatgacga cgataaggga
     2161 tccagcgcct ggtcccaccc tcagtttgag aagggcggag gctctggcgg cggaagcgga
     2221 ggatctgctt ggagccaccc ccagttcgaa aagtgatgag ggcccgttta aacccgctga
     2281 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2341 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2401 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2461 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2521 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2761 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2821 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2881 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2941 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3241 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3301 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3361 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3421 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3481 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3541 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3601 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3661 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3721 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3781 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3841 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3901 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3961 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4021 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4081 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4141 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4201 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4261 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4321 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4381 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4441 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4501 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5461 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5521 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5581 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5641 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5701 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5761 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5821 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5881 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5941 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6001 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6061 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6121 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6181 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6241 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6301 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6361 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6421 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6481 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6541 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6601 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6661 cacatttccc cgaaaagtgc cacctgacgt c
//
