LOCUS       V046920                 8255 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046920
VERSION     V046920
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..2094
                     /label="SQSTM1(NM_003900)"
                     /note="SQSTM1(NM_003900)"
                     /gene="SQSTM1"
     CDS             2095..2121
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2125..2151
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2155..2181
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2197..2253
                     /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             2254..2964
                     /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    3031..3619
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3685..3733
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3935..4363
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4377..4706
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4773..5567
                     /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    5743..5864
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5913..5929)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5937..5953
                     /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(5961..5991)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6006..6027
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6315..6903)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7074..7934)
                     /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(7935..8039)
                     /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 caccatggcg
      781 tcgctcaccg tgaaggccta ccttctgggc aaggaggacg cggcgcgcga gattcgccgc
      841 ttcagcttct gctgcagccc cgagcctgag gcggaagccg aggctgcggc gggtccggga
      901 ccctgcgagc ggctgctgag ccgggtggcc gccctgttcc ccgcgctgcg gcctggcggc
      961 ttccaggcgc actaccgcga tgaggacggg gacttggttg ccttttccag tgacgaggaa
     1021 ttgacaatgg ccatgtccta cgtgaaggat gacatcttcc gaatctacat taaagagaaa
     1081 aaagagtgcc ggcgggacca ccgcccaccg tgtgctcagg aggcgccccg caacatggtg
     1141 caccccaatg tgatctgcga tggctgcaat gggcctgtgg taggaacccg ctacaagtgc
     1201 agcgtctgcc cagactacga cttgtgtagc gtctgcgagg gaaagggctt gcaccggggg
     1261 cacaccaagc tcgcattccc cagccccttc gggcacctgt ctgagggctt ctcgcacagc
     1321 cgctggctcc ggaaggtgaa acacggacac ttcgggtggc caggatggga aatgggtcca
     1381 ccaggaaact ggagcccacg tcctcctcgt gcaggggagg cccgccctgg ccccacggca
     1441 gaatcagctt ctggtccatc ggaggatccg agtgtgaatt tcctgaagaa cgttggggag
     1501 agtgtggcag ctgcccttag ccctctgggc attgaagttg atatcgatgt ggagcacgga
     1561 gggaaaagaa gccgcctgac ccccgtctct ccagagagtt ccagcacaga ggagaagagc
     1621 agctcacagc caagcagctg ctgctctgac cccagcaagc cgggtgggaa tgttgagggc
     1681 gccacgcagt ctctggcgga gcagatgagg aagatcgcct tggagtccga ggggcgccct
     1741 gaggaacaga tggagtcgga taactgttca ggaggagatg atgactggac ccatctgtct
     1801 tcaaaagaag tggacccgtc tacaggtgaa ctccagtccc tacagatgcc agaatccgaa
     1861 gggccaagct ctctggaccc ctcccaggag ggacccacag ggctgaagga agctgccttg
     1921 tacccacatc tcccgccaga ggctgacccg cggctgattg agtccctctc ccagatgctg
     1981 tccatgggct tctctgatga aggcggctgg ctcaccaggc tcctgcagac caagaactat
     2041 gacatcggag cggctctgga caccatccag tattcaaagc atcccccgcc gttgtaccca
     2101 tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc tagctatcct
     2161 tatgatgtcc cagactacgc agaattcgga agcggagcta ctaacttcag cctgctgaag
     2221 caggctggcg acgtggagga gaaccctgga cctatggtga gcaagggcga ggaggataac
     2281 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2341 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2401 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2461 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2521 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2581 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2641 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2701 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2761 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2821 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2881 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2941 ggcatggacg agctgtacaa gtaactcgag tctagagggc cccttaaggg ttcgatccct
     3001 accggttagt aatgagtttg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3061 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3121 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3181 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3241 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3301 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3361 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3421 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3481 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3541 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3601 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     3661 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     3721 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3781 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3841 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3901 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3961 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4021 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4081 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4141 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4201 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4261 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4321 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4381 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4441 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4501 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4561 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4621 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4681 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4741 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4801 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4861 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4921 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4981 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5041 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5101 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5161 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5221 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5281 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5341 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5401 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5461 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5521 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5581 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5641 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5701 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5761 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5821 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5881 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6481 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6721 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8041 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8101 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8161 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8221 gcttgaccga caattgcatg aagaatctgc ttagg
//
