LOCUS       V046786                 9783 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046786
VERSION     V046786
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             776..2653
                     /label="MAG(NM_002361)"
                     /note="MAG(NM_002361)"
                     /gene="MAG"
     CDS             2654..2710
                     /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             3728..3781
                     /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&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             3782..4492
                     /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    4559..5147
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5213..5261
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5463..5891
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5905..6234
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6301..7095
                     /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    7271..7392
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7441..7457)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7465..7481
                     /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(7489..7519)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7534..7555
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7843..8431)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8602..9462)
                     /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(9463..9567)
                     /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 tcggatcccg ccaccatgat
      781 attcctcacg gcactgcctc tgttctggat tatgatttca gcctcccgag ggggtcactg
      841 gggtgcctgg atgccctcgt ccatctcggc cttcgaaggc acgtgcgtct ccatcccctg
      901 ccgctttgac ttcccggatg agctgcggcc cgctgtggtg catggtgtct ggtacttcaa
      961 tagcccctac cccaagaact accccccggt ggtcttcaag tcgcgcaccc aagtagtcca
     1021 cgagagcttc cagggccgca gccgcctcct gggggacctg ggcctgcgaa actgcaccct
     1081 cctgctcagc aacgtcagcc ccgagctggg cgggaagtac tacttccgtg gggacctggg
     1141 cggctacaac cagtacacct tctcagagca cagcgtcctg gatatcgtca acacccccaa
     1201 catcgtggtg cccccagagg tggtggcagg cacggaggtg gaggtcagct gcatggtgcc
     1261 ggacaactgc ccagagctgc gccctgagct gagctggctg ggccacgagg ggctggggga
     1321 gcccgctgtg ctgggccggc tgcgggagga cgagggcacc tgggtgcagg tgtcactgct
     1381 gcacttcgtg cccacgaggg aggccaacgg ccacaggctg ggctgccagg cctccttccc
     1441 caacaccacc ctgcagttcg agggctacgc cagcatggac gtcaagtacc ccccggtgat
     1501 tgtggagatg aactcctcgg tggaggccat cgagggctcc cacgtgagcc tgctctgtgg
     1561 ggctgacagc aaccccccgc cgctgctgac ctggatgcgg gacgggacag tcctccggga
     1621 ggcggtggcc gagagcctgc tcctggagct ggaggaggtg acccccgccg aagacggcgt
     1681 ctatgcctgc ctggccgaga atgcctatgg ccaggacaac cgcaccgtgg ggctcagtgt
     1741 catgtatgca ccctggaagc caacagtgaa cgggacaatg gtggccgtag agggggagac
     1801 ggtctctatc ttgtgctcca cacagagcaa cccggaccct attctcacca tcttcaagga
     1861 gaagcagatc ctgtccacgg tcatctacga gagcgagctg cagctggagc tgccggccgt
     1921 gtcacccgag gatgatggag agtactggtg tgtggctgag aaccagtatg gccagagggc
     1981 caccgccttc aacctgtctg tggagttcgc ccctgtgctc ctcctggagt cccactgcgc
     2041 ggcagcccga gacacggtgc agtgcctgtg cgtggtgaag tccaacccgg agccgtccgt
     2101 ggcctttgag ctgccatcgc gcaatgtgac cgtgaacgag agcgagcggg agttcgtgta
     2161 ctcggagcgc agcggcctcg tgctcaccag catcctcacg ctgcgggggc aggcccaggc
     2221 cccgccccgc gtcatctgca ccgcgaggaa cctctatggc gccaagagcc tggagctgcc
     2281 cttccaggga gcccatcgac tgatgtgggc caagatcggg cctgtgggcg ccgtggtcgc
     2341 ctttgccatc ctgattgcca tcgtctgcta cattacccag acacgcagga aaaagaacgt
     2401 gacagagagc cccagcttct cggcagggga caaccctccc gtcctgttca gcagcgactt
     2461 ccgcatctct ggggcaccag agaagtacga gagcgagagg cgcctgggat ctgagaggag
     2521 gctgctgggc cttcggggtg agcccccaga gctggacctg agctattctc actcggacct
     2581 ggggaaacgg cccaccaagg acagctacac gctgacggag gagctagctg agtatgctga
     2641 aatccgggtc aaggcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
     2701 tcctggaccg atgccgaaga gacgggacat cctagcgatc gtcctcatcg tgctgccctg
     2761 gactctgctc atcactgtct ggcaccagag caccctcgca cccctgctcg cggtacataa
     2821 ggatgagggc agtgaccccc gacgcgaaac gccgcccggc gccgacccca gggagtactg
     2881 cacgtctgac cgcgacatcg tggaggtggt gcgcaccgag tacgtgtaca cgcggccccc
     2941 gccatggtcc gacacgctgc ccaccatcca cgtggtgacg cccacctaca gccgcccggt
     3001 gcagaaggcc gagctgacgc gcatggccaa cacgctgctg cacgtgccca acctccactg
     3061 gctggtggtg gaggatgcgc cgcgccggac gccgctgacc gcgcgcctgc tgcgcgacac
     3121 cggcctcaac tacacgcacc tgcacgtgga gacgccccgc aactacaagc tgcgcggaga
     3181 cgcccgcgac ccacgcatcc cgcggggcac catgcagcgc aacctggccc tgcgctggct
     3241 gcgcgagacc ttcccgcgca actccagcca gcctggcgtg gtctacttcg ccgacgacga
     3301 caacacctac agcctggagc tcttcgaaga gatgcgcagc accaggaggg tgtccgtgtg
     3361 gcccgtcgcc ttcgtgggtg gcctgcggta cgaggcccca cgggtgaacg gggcagggaa
     3421 ggtggtcggc tggaagacgg tgtttgaccc ccaccggcca tttgcaatag acatggctgg
     3481 atttgccgtc aacctgcggc tcattctgca gcgaagccag gcctacttca agctgcgagg
     3541 tgtgaaggga ggctaccagg aaagcagcct ccttcgagaa cttgtcaccc tcaacgacct
     3601 ggagcccaag gcagccaact gcaccaagat cctggtgtgg cacacacgga cagagaagcc
     3661 agtgctggtg aatgagggca agaagggctt cactgacccc tcggtggaga tcgaattcgg
     3721 aagcggagag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     3781 tatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     3841 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3901 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     3961 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4021 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     4081 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4141 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     4201 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4261 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4321 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     4381 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     4441 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aactcgagtc
     4501 tagagggccc cttaagggtt cgatccctac cggttagtaa tgagtttgat atctcgacaa
     4561 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4621 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4681 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4741 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4801 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4861 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4921 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4981 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5041 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5101 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     5161 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5221 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5281 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5341 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5401 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     5461 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5521 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     5641 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5761 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5821 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5881 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     6001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6061 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     6181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6241 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     6301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6481 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6841 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     7021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7081 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     7141 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     7201 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     7261 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7321 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7381 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     7441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     8041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8281 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8581 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8641 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8701 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8761 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8821 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8881 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8941 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9001 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9061 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9121 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9181 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9241 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9301 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9361 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9421 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9481 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9541 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9601 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     9661 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9721 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9781 agg
//
