LOCUS       V040659                 8374 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040659
VERSION     V040659
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..3810
                     /label="GABBR1(NM_001470)"
                     /note="GABBR1(NM_001470)"
                     /gene="GABBR1"
     CDS             3817..3840
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             3847..3936
                     /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    3974..4198
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4244..4672
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4686..5015
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5082..5876
                     /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    6050..6171
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6220..6236)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6244..6260
                     /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(6268..6298)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6313..6334
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6622..7207)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7378..8238)
                     /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(8239..8343)
                     /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 ttgctgctgc tgctactggc gccactcttc
      961 ctccgccccc cgggcgcggg cggggcgcag acccccaacg ccacctcaga aggttgccag
     1021 atcatacacc cgccctggga agggggcatc aggtaccggg gcctgactcg ggaccaggtg
     1081 aaggctatca acttcctgcc agtggactat gagattgagt atgtgtgccg gggggagcgc
     1141 gaggtggtgg ggcccaaggt ccgcaagtgc ctggccaacg gctcctggac agatatggac
     1201 acacccagcc gctgtgtccg aatctgctcc aagtcttatt tgaccctgga aaatgggaag
     1261 gttttcctga cgggtgggga cctcccagct ctggacggag cccgggtgga tttccggtgt
     1321 gaccccgact tccatctggt gggcagctcc cggagcatct gtagtcaggg ccagtggagc
     1381 acccccaagc cccactgcca ggtgaatcga acgccacact cagaacggcg cgcagtgtac
     1441 atcggggcac tgtttcccat gagcgggggc tggccagggg gccaggcctg ccagcccgcg
     1501 gtggagatgg cgctggagga cgtgaatagc cgcagggaca tcctgccgga ctatgagctc
     1561 aagctcatcc accacgacag caagtgtgat ccaggccaag ccaccaagta cctatatgag
     1621 ctgctctaca acgaccctat caagatcatc cttatgcctg gctgcagctc tgtctccacg
     1681 ctggtggctg aggctgctag gatgtggaac ctcattgtgc tttcctatgg ctccagctca
     1741 ccagccctgt caaaccggca gcgtttcccc actttcttcc gaacgcaccc atcagccaca
     1801 ctccacaacc ctacccgcgt gaaactcttt gaaaagtggg gctggaagaa gattgctacc
     1861 atccagcaga ccactgaggt cttcacttcg actctggacg acctggagga acgagtgaag
     1921 gaggctggaa ttgagattac tttccgccag agtttcttct cagatccagc tgtgcccgtc
     1981 aaaaacctga agcgccagga tgcccgaatc atcgtgggac ttttctatga gactgaagcc
     2041 cggaaagttt tttgtgaggt gtacaaggag cgtctctttg ggaagaagta cgtctggttc
     2101 ctcattgggt ggtatgctga caattggttc aagatctacg acccttctat caactgcaca
     2161 gtggatgaga tgactgaggc ggtggagggc cacatcacaa ctgagattgt catgctgaat
     2221 cctgccaata cccgcagcat ttccaacatg acatcccagg aatttgtgga gaaactaacc
     2281 aagcgactga aaagacaccc tgaggagaca ggaggcttcc aggaggcacc gctggcctat
     2341 gatgccatct gggccttggc actggccctg aacaagacat ctggaggagg cggccgttct
     2401 ggtgtgcgcc tggaggactt caactacaac aaccagacca ttaccgacca aatctaccgg
     2461 gcaatgaact cttcgtcctt tgagggtgtc tctggccatg tggtgtttga tgccagcggc
     2521 tctcggatgg catggacgct tatcgagcag cttcagggtg gcagctacaa gaagattggc
     2581 tactatgaca gcaccaagga tgatctttcc tggtccaaaa cagataaatg gattggaggg
     2641 tcccccccag ctgaccagac cctggtcatc aagacattcc gcttcctgtc acagaaactc
     2701 tttatctccg tctcagttct ctccagcctg ggcattgtcc tagctgttgt ctgtctgtcc
     2761 tttaacatct acaactcaca tgtccgttat atccagaact cacagcccaa cctgaacaac
     2821 ctgactgctg tgggctgctc actggcttta gctgctgtct tccccctggg gctcgatggt
     2881 taccacattg ggaggaacca gtttcctttc gtctgccagg cccgcctctg gctcctgggc
     2941 ctgggcttta gtctgggcta cggttccatg ttcaccaaga tttggtgggt ccacacggtc
     3001 ttcacaaaga aggaagaaaa gaaggagtgg aggaagactc tggaaccctg gaagctgtat
     3061 gccacagtgg gcctgctggt gggcatggat gtcctcactc tcgccatctg gcagatcgtg
     3121 gaccctctgc accggaccat tgagacattt gccaaggagg aacctaagga agatattgac
     3181 gtctctattc tgccccagct ggagcattgc agctccagga agatgaatac atggcttggc
     3241 attttctatg gttacaaggg gctgctgctg ctgctgggaa tcttccttgc ttatgagacc
     3301 aagagtgtgt ccactgagaa gatcaatgat caccgggctg tgggcatggc tatctacaat
     3361 gtggcagtcc tgtgcctcat cactgctcct gtcaccatga ttctgtccag ccagcaggat
     3421 gcagcctttg cctttgcctc tcttgccata gttttctcct cctatatcac tcttgttgtg
     3481 ctctttgtgc ccaagatgcg caggctgatc acccgagggg aatggcagtc ggaggcgcag
     3541 gacaccatga agacagggtc atcgaccaac aacaacgagg aggagaagtc ccggctgttg
     3601 gagaaggaga accgtgaact ggaaaagatc attgctgaga aagaggagcg tgtctctgaa
     3661 ctgcgccatc aactccagtc tcggcagcag ctccgctccc ggcgccaccc accgacaccc
     3721 ccagaaccct ctgggggcct gcccagggga ccccctgagc cccccgaccg gcttagctgt
     3781 gatgggagtc gagtgcattt gctttataag ctcgaggatt acaaggatga cgacgataag
     3841 ggatccagcg cctggtccca ccctcagttt gagaagggcg gaggctctgg cggcggaagc
     3901 ggaggatctg cttggagcca cccccagttc gaaaagtgat gagggcccgt ttaaacccgc
     3961 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4021 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4081 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4141 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4201 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4261 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4321 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4381 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4441 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4501 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4561 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4621 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4681 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4741 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4801 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4861 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4921 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4981 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5041 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5101 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5161 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5221 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5281 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5341 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5401 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5461 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5521 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5581 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5641 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5701 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5761 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5821 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5881 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5941 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6001 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6181 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6241 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6301 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6361 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6421 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6481 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6961 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7021 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7081 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7141 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8341 gcgcacattt ccccgaaaag tgccacctga cgtc
//
