LOCUS       V028415                 8264 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028415
VERSION     V028415
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             941..3754
                     /label="GRIN1(NM_007327)"
                     /note="GRIN1(NM_007327)"
                     /gene="GRIN1"
     CDS             3761..3826
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3864..4088
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4134..4562
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4576..4905
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4972..5766
                     /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    5940..6061
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6110..6126)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6134..6150
                     /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(6158..6188)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6203..6224
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6512..7097)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7268..8128)
                     /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(8129..8233)
                     /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 aagcttggta ccgagctcgg atccgccacc atgagcacca tgcgcctgct
      961 gacgctcgcc ctgctgttct cctgctccgt cgcccgtgcc gcgtgcgacc ccaagatcgt
     1021 caacattggc gcggtgctga gcacgcggaa gcacgagcag atgttccgcg aggccgtgaa
     1081 ccaggccaac aagcggcacg gctcctggaa gattcagctc aatgccacct ccgtcacgca
     1141 caagcccaac gccatccaga tggctctgtc ggtgtgcgag gacctcatct ccagccaggt
     1201 ctacgccatc ctagttagcc atccacctac ccccaacgac cacttcactc ccacccctgt
     1261 ctcctacaca gccggcttct accgcatacc cgtgctgggg ctgaccaccc gcatgtccat
     1321 ctactcggac aagagcatcc acctgagctt cctgcgcacc gtgccgccct actcccacca
     1381 gtccagcgtg tggtttgaga tgatgcgtgt ctacagctgg aaccacatca tcctgctggt
     1441 cagcgacgac cacgagggcc gggcggctca gaaacgcctg gagacgctgc tggaggagcg
     1501 tgagtccaag gcagagaagg tgctgcagtt tgacccaggg accaagaacg tgacggccct
     1561 gctgatggag gcgaaagagc tggaggcccg ggtcatcatc ctttctgcca gcgaggacga
     1621 tgctgccact gtataccgcg cagccgcgat gctgaacatg acgggctccg ggtacgtgtg
     1681 gctggtcggc gagcgcgaga tctcggggaa cgccctgcgc tacgccccag acggcatcct
     1741 cgggctgcag ctcatcaacg gcaagaacga gtcggcccac atcagcgacg ccgtgggcgt
     1801 ggtggcccag gccgtgcacg agctcctcga gaaggagaac atcaccgacc cgccgcgggg
     1861 ctgcgtgggc aacaccaaca tctggaagac cgggccgctc ttcaagagag tgctgatgtc
     1921 ttccaagtat gcggatgggg tgactggtcg cgtggagttc aatgaggatg gggaccggaa
     1981 gttcgccaac tacagcatca tgaacctgca gaaccgcaag ctggtgcaag tgggcatcta
     2041 caatggcacc cacgtcatcc ctaatgacag gaagatcatc tggccaggcg gagagacaga
     2101 gaagcctcga gggtaccaga tgtccaccag actgaagatt gtgacgatcc accaggagcc
     2161 cttcgtgtac gtcaagccca cgctgagtga tgggacatgc aaggaggagt tcacagtcaa
     2221 cggcgaccca gtcaagaagg tgatctgcac cgggcccaac gacacgtcgc cgggcagccc
     2281 ccgccacacg gtgcctcagt gttgctacgg cttttgcatc gacctgctca tcaagctggc
     2341 acggaccatg aacttcacct acgaggtgca cctggtggca gatggcaagt tcggcacaca
     2401 ggagcgggtg aacaacagca acaagaagga gtggaatggg atgatgggcg agctgctcag
     2461 cgggcaggca gacatgatcg tggcgccgct aaccataaac aacgagcgcg cgcagtacat
     2521 cgagttttcc aagcccttca agtaccaggg cctgactatt ctggtcaaga aggagattcc
     2581 ccggagcacg ctggactcgt tcatgcagcc gttccagagc acactgtggc tgctggtggg
     2641 gctgtcggtg cacgtggtgg ccgtgatgct gtacctgctg gaccgcttca gccccttcgg
     2701 ccggttcaag gtgaacagcg aggaggagga ggaggacgca ctgaccctgt cctcggccat
     2761 gtggttctcc tggggcgtcc tgctcaactc cggcatcggg gaaggcgccc ccagaagctt
     2821 ctcagcgcgc atcctgggca tggtgtgggc cggctttgcc atgatcatcg tggcctccta
     2881 caccgccaac ctggcggcct tcctggtgct ggaccggccg gaggagcgca tcacgggcat
     2941 caacgaccct cggctgagga acccctcgga caagtttatc tacgccacgg tgaagcagag
     3001 ctccgtggat atctacttcc ggcgccaggt ggagctgagc accatgtacc ggcatatgga
     3061 gaagcacaac tacgagagtg cggcggaggc catccaggcc gtgagagaca acaagctgca
     3121 tgccttcatc tgggactcgg cggtgctgga gttcgaggcc tcgcagaagt gcgacctggt
     3181 gacgactgga gagctgtttt tccgctcggg cttcggcata ggcatgcgca aagacagccc
     3241 ctggaagcag aacgtctccc tgtccatcct caagtcccac gagaatggct tcatggaaga
     3301 cctggacaag acgtgggttc ggtatcagga atgtgactcg cgcagcaacg cccctgcgac
     3361 ccttactttt gagaacatgg ccggggtctt catgctggta gctgggggca tcgtggccgg
     3421 gatcttcctg attttcatcg agattgccta caagcggcac aaggatgctc gccggaagca
     3481 gatgcagctg gcctttgccg ccgttaacgt gtggcggaag aacctgcagg atagaaagag
     3541 tggtagagca gagcctgacc ctaaaaagaa agccacattt agggctatca cctccaccct
     3601 ggcttccagc ttcaagaggc gtaggtcctc caaagacacg agcaccgggg gtggacgcgg
     3661 cgctttgcaa aaccaaaaag acacagtgct gccgcgacgc gctattgaga gggaggaggg
     3721 ccagctgcag ctgtgttccc gtcataggga gagcctcgag gactacaaag accatgacgg
     3781 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     3841 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3901 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3961 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4021 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4081 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     4141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4321 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4441 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4501 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4561 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4621 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4681 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4741 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4801 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4861 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4921 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4981 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5041 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5101 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5161 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5221 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5281 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5341 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5401 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5461 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5521 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5581 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5641 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5701 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5761 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5821 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5881 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5941 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6001 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6061 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6121 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6181 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6241 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6301 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6361 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7021 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     7081 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7501 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7561 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7621 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7681 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7741 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7801 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7861 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7921 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7981 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8041 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8101 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8161 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8221 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
