LOCUS       V028548                 9655 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028548
VERSION     V028548
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2815..4302
                     /label="NOTUM(NM_178493)"
                     /note="NOTUM(NM_178493)"
                     /gene="NOTUM"
     CDS             4303..4320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    4358..4944
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             4964..5563
                     /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"
     misc_feature    5577..6165
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6372..7005
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7133..7149)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7157..7173
                     /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(7181..7211)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7226..7247
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7535..8123)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8294..9154)
                     /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(9155..9259)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9307..9441
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatgggc
     2821 cgaggggtgc gcgtgctgct gctgctgagc ctgctgcact gcgccggggg cagcgagggc
     2881 aggaagacct ggcggcgccg gggtcagcag ccgcctcctc ccccgcggac cgaggcggcg
     2941 ccggcggccg gacagcccgt ggagagcttc ccgctggact tcacggccgt ggagggtaac
     3001 atggacagct tcatggcgca agtcaagagc ctggcgcagt ccctgtaccc ctgctccgcg
     3061 cagcagctca acgaggacct gcgcctgcac ctcctactca acacctcggt gacctgcaac
     3121 gacggcagcc ccgccggcta ctacctgaag gagtccaggg gcagccggcg gtggctcctc
     3181 ttcctggaag gcggctggta ctgcttcaac cgcgagaact gcgactccag atacgacacc
     3241 atgcggcgcc tcatgagctc ccgggactgg ccgcgcactc gcacaggcac agggatcctg
     3301 tcctcacagc cggaggagaa cccctactgg tggaacgcaa acatggtctt catcccctac
     3361 tgctccagtg atgtttggag cggggcttca tccaagtctg agaagaacga gtacgccttc
     3421 atgggcgccc tcatcatcca ggaggtggtg cgggagcttc tgggcagagg gctgagcggg
     3481 gccaaggtgc tgctgctggc cgggagcagc gcggggggca ccggggtgct cctgaatgtg
     3541 gaccgtgtgg ctgagcagct ggagaagctg ggctacccag ccatccaggt gcgaggcctg
     3601 gctgactccg gctggttcct ggacaacaag cagtatcgcc acacagactg cgtcgacacg
     3661 atcacgtgcg cgcccacgga ggccatccgc cgtggcatca ggtactggaa cggggtggtc
     3721 ccggagcgct gccgacgcca gttccaggag ggcgaggagt ggaactgctt ctttggctac
     3781 aaggtctacc cgaccctgcg ctgccctgtg ttcgtggtgc agtggctgtt tgacgaggca
     3841 cagctgacgg tggacaacgt gcacctgacg gggcagccgg tgcaggaggg cctgcggctg
     3901 tacatccaga acctcggccg cgagctgcgc cacacactca aggacgtgcc ggccagcttt
     3961 gcccccgcct gcctctccca tgagatcatc atccggagcc actggacgga tgtccaggtg
     4021 aaggggacgt cgctgccccg agcactgcac tgctgggaca ggagcctcca tgacagccac
     4081 aaggccagca agacccccct caagggctgc cccgtccacc tggtggacag ctgcccctgg
     4141 ccccactgca acccctcatg ccccaccgtc cgagaccagt tcacggggca agagatgaac
     4201 gtggcccagt tcctcatgca catgggcttc gacatgcaga cggtggccca gccgcaggga
     4261 ctggagccca gtgagctgct ggggatgctg agcaacggaa gccatcatca ccatcaccat
     4321 tagctcgaga ctagttctag agcggccgcg gatcccgccc ctctccctcc ccccccccta
     4381 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     4441 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     4501 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     4561 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     4621 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     4681 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     4741 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     4801 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     4861 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     4921 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc
     4981 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     5041 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     5101 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5161 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5221 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5281 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5341 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5401 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5461 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5521 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc
     5581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5941 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     6001 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     6061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     6121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg
     6181 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     6241 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     6301 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6361 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     6421 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
     6481 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
     6541 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
     6601 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
     6661 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
     6721 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
     6781 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
     6841 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6901 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6961 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     7021 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt
     7081 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8341 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8401 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8461 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8521 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8581 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8641 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8701 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8761 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8821 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8881 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8941 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9001 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9061 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9121 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9181 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9241 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9301 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9361 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9421 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt
     9481 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt
     9541 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
     9601 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
