LOCUS       V027495                 9485 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027495
VERSION     V027495
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             775..3411
                     /label="GRM3(NM_000840)"
                     /note="GRM3(NM_000840)"
                     /gene="GRM3"
     CDS             3427..3483
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3484..4194
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    4261..4849
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4915..4963
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5165..5593
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5607..5936
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6003..6797
                     /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    6973..7094
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7143..7159)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7167..7183
                     /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(7191..7221)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7236..7257
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7545..8133)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8304..9164)
                     /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(9165..9269)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgaag
      781 atgttgacaa gactgcaagt tcttacctta gctttgtttt caaagggatt tttactctct
      841 ttaggggacc ataactttct aaggagagag attaaaatag aaggtgacct tgttttaggg
      901 ggcctgtttc ctattaacga aaaaggcact ggaactgaag aatgtgggcg aatcaatgaa
      961 gaccgaggga ttcaacgcct ggaagccatg ttgtttgcta ttgatgaaat caacaaagat
     1021 gattacttgc taccaggagt gaagttgggt gttcacattt tggatacatg ttcaagggat
     1081 acctatgcat tggagcaatc actggagttt gtcagggcat ctttgacaaa agtggatgaa
     1141 gctgagtata tgtgtcctga tggatcctat gccattcaag aaaacatccc acttctcatt
     1201 gcaggggtca ttggtggctc ttatagcagt gtttccatac aggtggcaaa cctgctgcgg
     1261 ctcttccaga tccctcagat cagctacgca tccaccagcg ccaaactcag tgataagtcg
     1321 cgctatgatt actttgccag gaccgtgccc cccgacttct accaggccaa agccatggct
     1381 gagatcttgc gcttcttcaa ctggacctac gtgtccacag tagcctccga gggtgattac
     1441 ggggagacag ggatcgaggc cttcgagcag gaagcccgcc tgcgcaacat ctgcatcgct
     1501 acggcggaga aggtgggccg ctccaacatc cgcaagtcct acgacagcgt gatccgagaa
     1561 ctgttgcaga agcccaacgc gcgcgtcgtg gtcctcttca tgcgcagcga cgactcgcgg
     1621 gagctcattg cagccgccag ccgcgccaat gcctccttca cctgggtggc cagcgacggc
     1681 tggggcgcgc aggagagcat catcaagggc agcgagcatg tggcctacgg cgccatcacc
     1741 ctggagctgg cctcccagcc tgtccgccag ttcgaccgct acttccagag cctcaacccc
     1801 tacaacaacc accgcaaccc ctggttccgg gacttctggg agcaaaagtt tcagtgcagc
     1861 ctccagaaca aacgcaacca caggcgcgtc tgcgacaagc acctggccat cgacagcagc
     1921 aactacgagc aagagtccaa gatcatgttt gtggtgaacg cggtgtatgc catggcccac
     1981 gctttgcaca aaatgcagcg caccctctgt cccaacacta ccaagctttg tgatgctatg
     2041 aagatcctgg atgggaagaa gttgtacaag gattacttgc tgaaaatcaa cttcacggct
     2101 ccattcaacc caaataaaga tgcagatagc atagtcaagt ttgacacttt tggagatgga
     2161 atggggcgat acaacgtgtt caatttccaa aatgtaggtg gaaagtattc ctacttgaaa
     2221 gttggtcact gggcagaaac cttatcgcta gatgtcaact ctatccactg gtcccggaac
     2281 tcagtcccca cttcccagtg cagcgacccc tgtgccccca atgaaatgaa gaatatgcaa
     2341 ccaggggatg tctgctgctg gatttgcatc ccctgtgaac cctacgaata cctggctgat
     2401 gagtttacct gtatggattg tgggtctgga cagtggccca ctgcagacct aactggatgc
     2461 tatgaccttc ctgaggacta catcaggtgg gaagacgcct gggccattgg cccagtcacc
     2521 attgcctgtc tgggttttat gtgtacatgc atggttgtaa ctgtttttat caagcacaac
     2581 aacacaccct tggtcaaagc atcgggccga gaactctgct acatcttatt gtttggggtt
     2641 ggcctgtcat actgcatgac attcttcttc attgccaagc catcaccagt catctgtgca
     2701 ttgcgccgac tcgggctggg gagttccttc gctatctgtt actcagccct gctgaccaag
     2761 acaaactgca ttgcccgcat cttcgatggg gtcaagaatg gcgctcagag gccaaaattc
     2821 atcagcccca gttctcaggt tttcatctgc ctgggtctga tcctggtgca aattgtgatg
     2881 gtgtctgtgt ggctcatcct ggaggcccca ggcaccagga ggtataccct tgcagagaag
     2941 cgggaaacag tcatcctaaa atgcaatgtc aaagattcca gcatgttgat ctctcttacc
     3001 tacgatgtga tcctggtgat cttatgcact gtgtacgcct tcaaaacgcg gaagtgccca
     3061 gaaaatttca acgaagctaa gttcataggt tttaccatgt acaccacgtg catcatctgg
     3121 ttggccttcc tccctatatt ttatgtgaca tcaagtgact acagagtgca gacgacaacc
     3181 atgtgcatct ctgtcagcct gagtggcttt gtggtcttgg gctgtttgtt tgcacccaag
     3241 gttcacatca tcctgtttca accccagaag aatgttgtca cacacagact gcacctcaac
     3301 aggttcagtg tcagtggaac tgggaccaca tactctcagt cctctgcaag cacgtatgtg
     3361 ccaacggtgt gcaatgggcg ggaagtcctc gactccacca cctcatctct ggaattcgga
     3421 agcggagcta ctaacttcag cctgctgaag caggctggcg acgtggagga gaaccctgga
     3481 cctatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     3541 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     3601 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     3661 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     3721 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     3781 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     3841 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     3901 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     3961 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4021 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4081 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4141 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaactcgag
     4201 tctagagggc cccttaaggg ttcgatccct accggttagt aatgagtttg atatctcgac
     4261 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4321 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4381 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4441 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4501 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4561 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4621 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4681 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4741 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4801 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     4861 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4921 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4981 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5041 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5101 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     5161 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5221 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5281 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     5341 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5401 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5461 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5521 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5581 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5761 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5941 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     6001 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     6061 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6121 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6181 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6241 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6301 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6361 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6421 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6481 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6541 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6601 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6661 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6721 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6781 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     6841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6961 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7021 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7081 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     7141 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7201 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7261 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7321 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     7741 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7981 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     8161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     8281 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8341 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8401 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8461 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8521 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8581 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8641 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8701 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8761 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8821 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8881 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8941 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     9001 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     9061 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     9121 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     9181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     9241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     9301 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     9361 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     9421 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     9481 ttagg
//
