LOCUS       V029461                 9010 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029461
VERSION     V029461
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        131..334
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             404..1276
                     /label="ASGR1(NM_001671)"
                     /note="ASGR1(NM_001671)"
                     /gene="ASGR1"
     CDS             1343..2053
                     /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    2093..2210
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2259..2769
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             2791..3390
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     LTR             3518..3751
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    3823..3944
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      3984..4119
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(4140..4158)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4168..4184)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      4326..4781
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4807..4911
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4912..5772
                     /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"
     rep_origin      5943..6531
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6819..6840
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6855..6885
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6893..6909
                     /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"
     primer_bind     6917..6933
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        6954..6972
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        7000..7226
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             7227..7407
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    7454..7579
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    8072..8305
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             8490..8534
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg actcagatct cgagctcaag cttcgaagcc accatgacca aggagtatca
      421 agaccttcag catctggaca atgaggagag tgaccaccat cagctcagaa aagggccacc
      481 tcctccccag cccctcctgc agcgtctctg ctccggacct cgcctcctcc tgctctccct
      541 gggcctcagc ctcctgctgc ttgtggttgt ctgtgtgatc ggatcccaaa actcccagct
      601 gcaggaggag ctgcggggcc tgagagagac gttcagcaac ttcacagcga gcacggaggc
      661 ccaggtcaag ggcttgagca cccagggagg caatgtggga agaaagatga agtcgctaga
      721 gtcccagctg gagaaacagc agaaggacct gagtgaagat cactccagcc tgctgctcca
      781 cgtgaagcag ttcgtgtctg acctgcggag cctgagctgt cagatggcgg cgctccaggg
      841 caatggctca gaaaggacct gctgcccggt caactgggtg gagcacgagc gcagctgcta
      901 ctggttctct cgctccggga aggcctgggc tgacgccgac aactactgcc ggctggagga
      961 cgcgcacctg gtggtggtca cgtcctggga ggagcagaaa tttgtccagc accacatagg
     1021 ccctgtgaac acctggatgg gcctccacga ccaaaacggg ccctggaagt gggtggacgg
     1081 gacggactac gagacgggct tcaagaactg gaggccggag cagccggacg actggtacgg
     1141 ccacgggctc ggaggaggcg aggactgtgc ccacttcacc gacgacggcc gctggaacga
     1201 cgacgtctgc cagaggccct accgctgggt ctgcgagaca gagctggaca aggccagcca
     1261 ggagccacct ctccttggag gagggggaag cggaggcggt gggtcgacgg taccgcgggc
     1321 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat
     1381 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     1441 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     1501 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     1561 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     1621 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     1681 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     1741 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     1801 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     1861 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     1921 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     1981 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     2041 gctgtacaag tgaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa
     2101 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2161 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2221 acagggacag cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc
     2281 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
     2341 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
     2401 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
     2461 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
     2521 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg
     2581 gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg
     2641 gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc
     2701 tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc
     2761 tctccccagg gggatccacc ggagcttacc atgaccgagt acaagcccac ggtgcgcctc
     2821 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     2881 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     2941 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3001 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     3061 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     3121 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     3181 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     3241 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     3301 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     3361 tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc
     3421 gaaaggagcg cacgacccca tgcatcggta cctttaagac caatgactta caaggcagct
     3481 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3541 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     3601 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3661 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3721 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     3781 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     3841 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3901 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     3961 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4021 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4081 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     4141 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4201 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4261 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4321 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4381 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4441 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4501 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4561 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4621 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4681 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4741 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     4801 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4861 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4921 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4981 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5041 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5101 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5161 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5221 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5281 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5341 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5401 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5461 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5521 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5581 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5641 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5701 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5761 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5821 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5881 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5941 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6001 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6061 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6601 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6661 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6721 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6781 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6841 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6901 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     6961 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
     7021 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
     7081 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
     7141 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
     7201 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
     7261 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7321 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7381 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     7441 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     7501 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     7561 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     7621 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     7681 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     7741 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     7801 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     7861 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     7921 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     7981 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     8041 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     8101 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     8161 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     8221 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     8281 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     8341 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     8401 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     8461 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     8521 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     8581 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     8641 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     8701 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     8761 ccattcgatt agtgaacgga tctcgacggt atcgatcacg agactagcct cgagcggccg
     8821 cccccttcac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     8881 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     8941 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     9001 tggactatca
//
