LOCUS       V027594                 8501 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027594
VERSION     V027594
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..2427
                     /label="GLDN(NM_181789)"
                     /note="GLDN(NM_181789)"
                     /gene="GLDN"
     CDS             2443..2499
                     /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             2500..3210
                     /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    3277..3865
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3931..3979
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4181..4609
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4623..4952
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5019..5813
                     /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    5989..6110
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6159..6175)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6183..6199
                     /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(6207..6237)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6252..6273
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6561..7149)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7320..8180)
                     /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(8181..8285)
                     /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 caccatggcc
      781 cgaggcgctg agggaggccg tggggacgcg ggttggggcc tgcgtggcgc cctggcggcc
      841 gtggcgctgc tctcggcgct caacgctgcg ggcacggtgt tcgcgctgtg ccagtggcgc
      901 gggctgagct cggcgctgcg ggctttggag gcgcagcggg gccgggagca gcgcgaggac
      961 agtgccctgc gctccttcct ggccgagttg agccgcgcgc cgcgcggggc gtccgcacca
     1021 ccccaagacc cggccagctc agctcgcaac aagcgcagcc acagcggcga gcccgcgccg
     1081 catatccgcg ccgagagcca tgacatgctg atgatgatga cctactccat ggtgccgatc
     1141 cgagtgatgg tggacctgtg caacagcacc aagggcatct gcctcacagg accttctgga
     1201 ccaccaggac ctccgggagc cggcgggttg ccaggacaca acggattgga tggacagcct
     1261 ggtcctcagg gcccaaaagg agaaaaagga gcaaatggaa aaagaggaaa aatggggata
     1321 cctggagctg caggaaatcc aggggaaagg ggagaaaagg gagaccatgg tgaactgggc
     1381 ctgcagggaa atgagggccc accagggcag aagggagaaa agggtgacaa aggagatgtg
     1441 tccaacgacg tgctcctggc aggtgccaaa ggtgaccaag gcccacccgg tccacctggg
     1501 cccccaggcc ctccaggtcc tccagggccc cctggaagca gaagagccaa aggccctcgg
     1561 cagccaagca tgttcaacgg ccagtgccca ggtgagactt gtgccatacc aaatgatgat
     1621 accttggttg gaaaagctga tgagaaagcc agtgaacacc attccccaca agcagaatcc
     1681 atgatcactt ccattggaaa cccagtgcaa gtactgaaag tgacagagac atttgggact
     1741 tggataagag agtctgctaa caagagtgat gaccggattt gggtgacaga gcatttttca
     1801 ggcatcatgg ttaaggaatt caaggatcag ccctcacttc tgaatggcag ttacacgttc
     1861 atccaccttc catactattt ccatggctgt gggcacgttg tttacaacaa ctctctctac
     1921 taccacaaag ggggttctaa taccctagtg agatttgaat ttggccagga aacatcccaa
     1981 actctgaagc ttgaaaatgc cttgtatttt gatcgaaaat acctttttgc aaattccaaa
     2041 acttacttca atctagctgt agatgaaaag ggcctttgga ttatctatgc gtcaagtgtg
     2101 gacggctcga gcattcttgt agcacaactg gatgagagga cattctcagt ggtgcaacac
     2161 gtcaatacca cgtaccctaa atccaaggct ggcaacgcct tcattgcccg aggaatcctc
     2221 tatgtcacag acaccaaaga tatgagggtc acatttgcct ttgatttgtt aggagggaaa
     2281 cagatcaatg caaactttga tttaagaact tcccagtctg ttcttgccat gttagcatac
     2341 aacatgagag atcagcattt atattcatgg gaagatggcc atttaatgct ttatcctgtg
     2401 cagtttttgt caactacctt aaatcaggaa ttcggaagcg gagctactaa cttcagcctg
     2461 ctgaagcagg ctggcgacgt ggaggagaac cctggaccta tggtgagcaa gggcgaggag
     2521 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     2581 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2641 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2701 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2761 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2821 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2881 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2941 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3001 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3061 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     3121 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     3181 accggcggca tggacgagct gtacaagtaa ctcgagtcta gagggcccct taagggttcg
     3241 atccctaccg gttagtaatg agtttgatat ctcgacaatc aacctctgga ttacaaaatt
     3301 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3361 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3421 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3481 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3541 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3601 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3661 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     3721 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3781 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3841 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     3901 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     3961 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4021 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4081 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4141 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4321 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4381 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4501 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     4561 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4621 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4681 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4861 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4921 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4981 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5041 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5101 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5161 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5221 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5281 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5341 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5401 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5461 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5521 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5581 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5641 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5701 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5761 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5821 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5881 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5941 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     6001 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6061 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6121 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6181 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6241 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6301 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6361 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6421 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6481 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6541 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6601 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6661 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6721 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6781 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6841 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6901 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6961 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7021 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7081 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7141 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7201 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7261 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7321 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7381 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7441 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7501 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7561 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7621 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7681 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7741 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7801 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7861 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7921 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7981 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8041 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8101 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8161 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8221 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8281 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8341 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     8401 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     8461 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
