LOCUS       V041601                 8305 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041601
VERSION     V041601
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..3741
                     /label="GRIN1(NM_007327)"
                     /note="GRIN1(NM_007327)"
                     /gene="GRIN1"
     CDS             3748..3771
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             3778..3867
                     /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    3905..4129
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4175..4603
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4617..4946
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5013..5807
                     /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    5981..6102
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6151..6167)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6175..6191
                     /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(6199..6229)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6244..6265
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6553..7138)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7309..8169)
                     /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(8170..8274)
                     /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 agcaccatgc gcctgctgac gctcgccctg
      961 ctgttctcct gctccgtcgc ccgtgccgcg tgcgacccca agatcgtcaa cattggcgcg
     1021 gtgctgagca cgcggaagca cgagcagatg ttccgcgagg ccgtgaacca ggccaacaag
     1081 cggcacggct cctggaagat tcagctcaat gccacctccg tcacgcacaa gcccaacgcc
     1141 atccagatgg ctctgtcggt gtgcgaggac ctcatctcca gccaggtcta cgccatccta
     1201 gttagccatc cacctacccc caacgaccac ttcactccca cccctgtctc ctacacagcc
     1261 ggcttctacc gcatacccgt gctggggctg accacccgca tgtccatcta ctcggacaag
     1321 agcatccacc tgagcttcct gcgcaccgtg ccgccctact cccaccagtc cagcgtgtgg
     1381 tttgagatga tgcgtgtcta cagctggaac cacatcatcc tgctggtcag cgacgaccac
     1441 gagggccggg cggctcagaa acgcctggag acgctgctgg aggagcgtga gtccaaggca
     1501 gagaaggtgc tgcagtttga cccagggacc aagaacgtga cggccctgct gatggaggcg
     1561 aaagagctgg aggcccgggt catcatcctt tctgccagcg aggacgatgc tgccactgta
     1621 taccgcgcag ccgcgatgct gaacatgacg ggctccgggt acgtgtggct ggtcggcgag
     1681 cgcgagatct cggggaacgc cctgcgctac gccccagacg gcatcctcgg gctgcagctc
     1741 atcaacggca agaacgagtc ggcccacatc agcgacgccg tgggcgtggt ggcccaggcc
     1801 gtgcacgagc tcctcgagaa ggagaacatc accgacccgc cgcggggctg cgtgggcaac
     1861 accaacatct ggaagaccgg gccgctcttc aagagagtgc tgatgtcttc caagtatgcg
     1921 gatggggtga ctggtcgcgt ggagttcaat gaggatgggg accggaagtt cgccaactac
     1981 agcatcatga acctgcagaa ccgcaagctg gtgcaagtgg gcatctacaa tggcacccac
     2041 gtcatcccta atgacaggaa gatcatctgg ccaggcggag agacagagaa gcctcgaggg
     2101 taccagatgt ccaccagact gaagattgtg acgatccacc aggagccctt cgtgtacgtc
     2161 aagcccacgc tgagtgatgg gacatgcaag gaggagttca cagtcaacgg cgacccagtc
     2221 aagaaggtga tctgcaccgg gcccaacgac acgtcgccgg gcagcccccg ccacacggtg
     2281 cctcagtgtt gctacggctt ttgcatcgac ctgctcatca agctggcacg gaccatgaac
     2341 ttcacctacg aggtgcacct ggtggcagat ggcaagttcg gcacacagga gcgggtgaac
     2401 aacagcaaca agaaggagtg gaatgggatg atgggcgagc tgctcagcgg gcaggcagac
     2461 atgatcgtgg cgccgctaac cataaacaac gagcgcgcgc agtacatcga gttttccaag
     2521 cccttcaagt accagggcct gactattctg gtcaagaagg agattccccg gagcacgctg
     2581 gactcgttca tgcagccgtt ccagagcaca ctgtggctgc tggtggggct gtcggtgcac
     2641 gtggtggccg tgatgctgta cctgctggac cgcttcagcc ccttcggccg gttcaaggtg
     2701 aacagcgagg aggaggagga ggacgcactg accctgtcct cggccatgtg gttctcctgg
     2761 ggcgtcctgc tcaactccgg catcggggaa ggcgccccca gaagcttctc agcgcgcatc
     2821 ctgggcatgg tgtgggccgg ctttgccatg atcatcgtgg cctcctacac cgccaacctg
     2881 gcggccttcc tggtgctgga ccggccggag gagcgcatca cgggcatcaa cgaccctcgg
     2941 ctgaggaacc cctcggacaa gtttatctac gccacggtga agcagagctc cgtggatatc
     3001 tacttccggc gccaggtgga gctgagcacc atgtaccggc atatggagaa gcacaactac
     3061 gagagtgcgg cggaggccat ccaggccgtg agagacaaca agctgcatgc cttcatctgg
     3121 gactcggcgg tgctggagtt cgaggcctcg cagaagtgcg acctggtgac gactggagag
     3181 ctgtttttcc gctcgggctt cggcataggc atgcgcaaag acagcccctg gaagcagaac
     3241 gtctccctgt ccatcctcaa gtcccacgag aatggcttca tggaagacct ggacaagacg
     3301 tgggttcggt atcaggaatg tgactcgcgc agcaacgccc ctgcgaccct tacttttgag
     3361 aacatggccg gggtcttcat gctggtagct gggggcatcg tggccgggat cttcctgatt
     3421 ttcatcgaga ttgcctacaa gcggcacaag gatgctcgcc ggaagcagat gcagctggcc
     3481 tttgccgccg ttaacgtgtg gcggaagaac ctgcaggata gaaagagtgg tagagcagag
     3541 cctgacccta aaaagaaagc cacatttagg gctatcacct ccaccctggc ttccagcttc
     3601 aagaggcgta ggtcctccaa agacacgagc accgggggtg gacgcggcgc tttgcaaaac
     3661 caaaaagaca cagtgctgcc gcgacgcgct attgagaggg aggagggcca gctgcagctg
     3721 tgttcccgtc atagggagag cctcgaggat tacaaggatg acgacgataa gggatccagc
     3781 gcctggtccc accctcagtt tgagaagggc ggaggctctg gcggcggaag cggaggatct
     3841 gcttggagcc acccccagtt cgaaaagtga tgagggcccg tttaaacccg ctgatcagcc
     3901 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3961 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4021 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4081 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4141 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4201 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4261 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4321 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4381 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4441 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4501 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4561 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4621 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4681 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4801 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4861 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4921 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4981 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5041 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5101 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5161 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5221 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5281 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5341 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5401 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5461 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5521 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5581 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5641 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5701 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5761 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5821 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5881 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5941 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6061 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6121 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6181 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6241 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6361 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6421 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6481 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6541 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6601 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6661 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6721 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6781 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6841 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6901 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6961 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7021 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7081 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8281 tccccgaaaa gtgccacctg acgtc
//
