LOCUS       V031664                 8555 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031664
VERSION     V031664
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             757..2196
                     /label="HTRA1(NM_002775)"
                     /note="HTRA1(NM_002775)"
                     /gene="HTRA1"
     CDS             2497..2553
                     /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             2554..3264
                     /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    3331..3919
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3985..4033
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4235..4663
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4677..5006
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5073..5867
                     /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    6043..6164
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6213..6229)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6237..6253
                     /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(6261..6291)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6306..6327
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6615..7203)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7374..8234)
                     /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(8235..8339)
                     /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 gccaccatgc agatcccgcg cgccgctctt
      781 ctcccgctgc tgctgctgct gctggcggcg cccgcctcgg cgcagctgtc ccgggccggc
      841 cgctcggcgc ctttggccgc cgggtgccca gaccgctgcg agccggcgcg ctgcccgccg
      901 cagccggagc actgcgaggg cggccgggcc cgggacgcgt gcggctgctg cgaggtgtgc
      961 ggcgcgcccg agggcgccgc gtgcggcctg caggagggcc cgtgcggcga ggggctgcag
     1021 tgcgtggtgc ccttcggggt gccagcctcg gccacggtgc ggcggcgcgc gcaggccggc
     1081 ctctgtgtgt gcgccagcag cgagccggtg tgcggcagcg acgccaacac ctacgccaac
     1141 ctgtgccagc tgcgcgccgc cagccgccgc tccgagaggc tgcaccggcc gccggtcatc
     1201 gtcctgcagc gcggagcctg cggccaaggg caggaagatc ccaacagttt gcgccataaa
     1261 tataacttta tcgcggacgt ggtggagaag atcgcccctg ccgtggttca tatcgaattg
     1321 tttcgcaagc ttccgttttc taaacgagag gtgccggtgg ctagtgggtc tgggtttatt
     1381 gtgtcggaag atggactgat cgtgacaaat gcccacgtgg tgaccaacaa gcaccgggtc
     1441 aaagttgagc tgaagaacgg tgccacttac gaagccaaaa tcaaggatgt ggatgagaaa
     1501 gcagacatcg cactcatcaa aattgaccac cagggcaagc tgcctgtcct gctgcttggc
     1561 cgctcctcag agctgcggcc gggagagttc gtggtcgcca tcggaagccc gttttccctt
     1621 caaaacacag tcaccaccgg gatcgtgagc accacccagc gaggcggcaa agagctgggg
     1681 ctccgcaact cagacatgga ctacatccag accgacgcca tcatcaacta tggaaactcg
     1741 ggaggcccgt tagtaaacct ggacggtgaa gtgattggaa ttaacacttt gaaagtgaca
     1801 gctggaatct cctttgcaat cccatctgat aagattaaaa agttcctcac ggagtcccat
     1861 gaccgacagg ccaaaggaaa agccatcacc aagaagaagt atattggtat ccgaatgatg
     1921 tcactcacgt ccagcaaagc caaagagctg aaggaccggc accgggactt cccagacgtg
     1981 atctcaggag cgtatataat tgaagtaatt cctgataccc cagcagaagc tggtggtctc
     2041 aaggaaaacg acgtcataat cagcatcaat ggacagtccg tggtctccgc caatgatgtc
     2101 agcgacgtca ttaaaaggga aagcaccctg aacatggtgg tccgcagggg taatgaagat
     2161 atcatgatca cagtgattcc cgaagaaatt gacccagaat tcatgattct ctgggaaaag
     2221 aaaagcgaag atattaccga gacattcaac cccgacaaca acatgatcgc cgccgacgcg
     2281 agcgaggcat cggaggcggg gaaggcagaa aacacccatt acaccggcaa gcaccgcttc
     2341 atgtaccgga tcctgttcgt cctgacctgc ctgatcacct tcattgtggt ggctattgtc
     2401 ggcggaatta tcgcttcgaa cagaaacgtc ggaaatagaa tagctcaagg ccaaggatta
     2461 gccttgaaat tagcaggtct tgatatcgga agcggagcta ctaacttcag cctgctgaag
     2521 caggctggcg acgtggagga gaaccctgga cctatggtga gcaagggcga ggaggataac
     2581 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2641 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2701 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2761 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2821 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2881 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2941 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     3001 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     3061 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     3121 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     3181 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     3241 ggcatggacg agctgtacaa gtaactcgag tctagagggc cccttaaggg ttcgatccct
     3301 accggttagt aatgagtttg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3361 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3421 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3481 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3541 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3601 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3661 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3721 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3781 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3841 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3901 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     3961 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     4021 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4081 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4141 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     4201 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4261 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4321 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4381 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4441 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4501 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4561 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4621 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4681 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4741 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4801 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4861 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4921 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4981 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5041 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5101 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5161 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5221 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5281 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5341 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5401 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5461 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5521 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5581 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5641 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5701 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5761 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5821 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5881 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5941 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6001 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6061 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6121 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6181 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6241 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6301 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6361 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6421 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6781 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7021 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7381 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7501 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7681 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7741 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7801 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7861 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7921 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7981 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8041 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8101 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8161 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8221 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8281 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8341 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8401 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8461 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8521 gcttgaccga caattgcatg aagaatctgc ttagg
//
