LOCUS       V032194                 7490 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032194
VERSION     V032194
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             624..3437
                     /label="Grin1(NM_008169)"
                     /note="Grin1(NM_008169)"
                     /gene="Grin1"
     CDS             3480..4157
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="improved tetrameric variant of DsRed fluorescent
                     protein"
                     /transl_table=1
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL*"
                     /label="DsRed2"
     polyA_signal    4278..4399
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4406..4861)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4888..4992
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4994..5351
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5386..6180
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6412..6459
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6788..7376
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatgagca ccatgcacct gctgacattc gccctgcttt
      661 tctcctgctc cttcgcccgc gctgcctgcg accccaagat tgtcaacatc ggcgcggtgc
      721 tgagcacgcg caagcacgag cagatgttcc gcgaggcagt aaaccaggcc aataagcgac
      781 acggctcttg gaagatacag ctcaacgcca cttctgtcac ccacaagccc aacgccatac
      841 agatggccct gtcagtgtgt gaggacctca tctctagcca ggtctacgct atcctagtta
      901 gtcacccgcc tactcccaac gaccacttca ctcccacccc tgtctcctac acagctggct
      961 tctacagaat ccccgtcctg gggctgacta cccgaatgtc catctactct gacaagagca
     1021 tccacctgag cttccttcgc accgtaccac cctactccca ccagtccagc gtctggtttg
     1081 agatgatgcg cgtctacaac tggaaccata tcatcctgct ggtcagcgat gaccacgagg
     1141 gccgggcagc gcagaagcgc ctggagacgt tgctggagga gcgtgagtcc aaggcagaga
     1201 aggtgctgca gtttgaccca ggaaccaaga atgtgacggc tctgctgatg gaagcccggg
     1261 acctggaagc ccgggtcatc atcctttctg caagcgagga cgacgctgcc accgtatacc
     1321 gcgcagccgc gatgctgaac atgactggct ctgggtacgt gtggctcgtc ggggagcgcg
     1381 agatctctgg gaatgccctg cgctacgctc ctgacggcat catcggactt cagctaatca
     1441 acggcaagaa cgagtcggcc cacatcagtg acgctgtggg cgtggtggca caggcagtcc
     1501 acgagctcct agaaaaggag aacatcactg atccaccgcg gggttgcgtg ggcaacacca
     1561 acatctggaa gacaggacca ctgttcaaga gggtgctgat gtcttccaag tatgcagatg
     1621 gagtgactgg ccgtgtggaa ttcaatgagg atggggaccg gaagtttgcc aactatagta
     1681 tcatgaacct gcagaaccgc aagctggtgc aagtgggcat ctacaatggt acccatgtca
     1741 tcccaaatga caggaagatc atctggccag gaggagagac agagaagcct cgaggatacc
     1801 agatgtccac cagactaaag atagtgacaa tccaccaaga acccttcgtg tatgtcaagc
     1861 ccacaatgag tgatggcaca tgcaaagagg agttcacagt caatggtgac cctgtcaaga
     1921 aggtgatctg tacggggcct aatgacacat ccccaggaag cccacgtcac acagtgcccc
     1981 agtgctgtta tggcttctgc gttgacctgc tcatcaagct ggcacggacc atgaatttta
     2041 cctacgaggt gcaccttgtg gcagatggca agtttggcac acaggagcgg gtaaacaaca
     2101 gcaacaaaaa ggagtggaac ggaatgatgg gagagctgct cagtggtcaa gcagacatga
     2161 tcgtggctcc actgaccatt aacaatgagc gtgcgcagta catagagttc tccaagccct
     2221 tcaagtacca gggcctgacc attctggtca agaaggagat ccctcggagc acactggact
     2281 cattcatgca gccctttcag agcacactgt ggctgctggt ggggctgtca gttcatgtgg
     2341 tggccgtgat gctgtacctg ctggaccgct tcagtccctt tggccgattt aaggtgaaca
     2401 gcgaggagga ggaggaggat gcactgaccc tgtcctctgc catgtggttt tcctggggcg
     2461 tcctgctcaa ctctggcatt ggggaaggtg ccccccggag tttctctgct cgtatcctag
     2521 gcatggtgtg ggctggtttt gccatgatca tcgtggcttc ctacactgcc aacctggcag
     2581 ccttcctggt gctggatagg cctgaggagc gcatcacagg catcaatgac cccaggctca
     2641 gaaacccctc agacaagttc atctatgcaa ctgtaaaaca gagctctgtg gatatctact
     2701 tccggaggca ggtggagttg agcaccatgt accggcacat ggagaagcac aattatgaga
     2761 gtgcagctga ggccatccag gctgtgcggg acaacaagct ccatgccttc atctgggact
     2821 cagctgtgct ggagtttgag gcttcacaga agtgcgatct ggtgaccacg ggtgagctgt
     2881 tcttccgctc cggctttggc atcggcatgc gcaaggacag cccctggaag caaaatgtgt
     2941 ccctgtccat actcaagtcc catgagaatg gcttcatgga agacctggat aagacatggg
     3001 ttcggtatca agaatgtgac tcccgcagca atgcccctgc caccctcact tttgagaaca
     3061 tggcaggggt cttcatgctg gtggctggag gcatcgtagc tgggatcttc ctcattttca
     3121 tcgagatcgc ctacaagcga cacaaggatg cccgtaggaa gcagatgcag ctggcttttg
     3181 cagccgtgaa cgtgtggagg aagaacctgc aggatagaaa gagtggtaga gcagagcccg
     3241 accctaaaaa gaaagccaca tttagggcta tcacctccac cctggcctcc agcttcaaga
     3301 gacgtaggtc ctccaaagac acgagcaccg ggggtggacg cggcgctttg caaaaccaaa
     3361 aagacacagt gctgccgcga cgcgctattg agagggagga gggccagctg cagctgtgtt
     3421 cccgtcatag ggagagcgga ggtggaggtt caggtggagg tggatccccg gtcgccacca
     3481 tggcctcctc cgagaacgtc atcaccgagt tcatgcgctt caaggtgcgc atggagggca
     3541 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggcc
     3601 acaacaccgt gaagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     3661 tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg
     3721 actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3781 acggcggcgt ggcgaccgtg acccaggact cctccctgca ggacggctgc ttcatctaca
     3841 aggtgaagtt catcggcgtg aacttcccct ccgacggccc cgtgatgcag aagaagacca
     3901 tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga
     3961 cccacaaggc cctgaagctg aaggacggcg gccactacct ggtggagttc aagtccatct
     4021 acatggccaa gaagcccgtg cagctgcccg gctactacta cgtggacgcc aagctggaca
     4081 tcacctccca caacgaggac tacaccatcg tggagcagta cgagcgcacc gagggccgcc
     4141 accacctgtt cctgtagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4201 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4261 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4381 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4441 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4501 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4561 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4621 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4681 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4741 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4801 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4981 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     5041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5101 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5161 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5221 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5281 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5341 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5401 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5461 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5521 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5581 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5641 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5701 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5761 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5821 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5881 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5941 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6001 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6061 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6121 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6181 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6241 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6301 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6361 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6421 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6481 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6541 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6601 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6661 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6901 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7441 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
