LOCUS       V027520                 9405 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027520
VERSION     V027520
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2301..4259
                     /label="TRIM32(NM_012210)"
                     /note="TRIM32(NM_012210)"
                     /gene="TRIM32"
     CDS             4266..4331
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4375..4586
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4599..4867
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4894..5493
                     /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    5500..6088
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6162..6395
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6467..6588
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6607..6742
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6780..6796)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6804..6820
                     /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(6828..6858)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6873..6894
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7182..7770)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7941..8801)
                     /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(8802..8906)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9380..9396
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atggctgcag cagcagcttc tcacctgaac ctggatgccc
     2341 tccgggaagt gctagaatgc cccatctgca tggagtcctt cacagaagag cagctgcgtc
     2401 ccaagcttct gcactgtggc cataccatct gccgccagtg cctggagaag ctattggcca
     2461 gtagcatcaa tggtgtccgc tgtccctttt gcagcaagat tacccgcata accagcttga
     2521 cccagctgac agacaatctg acagtgctaa agatcattga tacagctggg ctcagcgagg
     2581 ctgtggggct gctcatgtgt cggtcctgtg ggcggcgtct gccccggcaa ttctgccgga
     2641 gctgtggttt ggtgttatgt gagccctgcc gggaggcaga ccatcagcct cctggccact
     2701 gtacactccc tgtcaaagaa gcagctgagg agcggcgtcg ggactttgga gagaagttaa
     2761 ctcgtctgcg ggaacttatg ggggagctgc agcggcggaa ggcagccttg gaaggtgtct
     2821 ccaaggacct tcaggcaagg tataaagcag ttctccagga gtatgggcat gaggagcgca
     2881 gggtccagga tgagctggct cgctctcgga agttcttcac aggctctttg gctgaagttg
     2941 agaagtccaa tagtcaagtg gtagaggagc agagttacct gcttaacatt gcagaggtgc
     3001 aggctgtgtc tcgctgtgac tacttcctgg ccaagatcaa gcaggcagat gtagcactac
     3061 tggaggagac agctgatgag gaggagccag agctcactgc cagcttgcct cgggagctca
     3121 ccctgcaaga tgtggagctc cttaaggtag gtcatgttgg ccccctccaa attggacaag
     3181 ctgttaagaa gccccggaca gttaacgtgg aagattcctg ggccatggag gccacagcgt
     3241 ctgctgcctc tacctctgtt acttttagag agatggacat gagcccggag gaagtggttg
     3301 ccagccctag ggcctcacct gctaaacagc ggggtcctga ggcagcctcc aatatccagc
     3361 agtgcctctt tctcaagaag atgggggcca aaggcagcac tccaggaatg ttcaatcttc
     3421 cagtcagtct ctacgtgacc agtcaaggtg aagtactagt cgctgaccgt ggtaactatc
     3481 gtatacaagt ctttacccgc aaaggctttt tgaaggaaat ccgccgcagc cccagtggca
     3541 ttgatagctt tgtgctaagc ttccttgggg cagatctacc caacctcact cctctctcag
     3601 tggcaatgaa ctgccagggg ctgattggtg tgactgacag ctatgataac tccctcaagg
     3661 tatatacctt ggatggccac tgcgtggcct gtcacaggag ccagctgagc aaaccatggg
     3721 gtatcacagc cttgccatct ggccagtttg tagtaaccga tgtggaaggt ggaaagcttt
     3781 ggtgtttcac agttgatcga ggatcagggg tggtcaaata cagctgccta tgtagtgctg
     3841 tgcggcccaa atttgtcacc tgtgatgctg agggcaccgt ctacttcacc cagggcttag
     3901 gcctcaatct ggagaatcgg cagaatgagc accacctgga gggtggcttt tccattggct
     3961 ctgtaggccc tgatgggcag ctgggtcgcc agattagcca cttcttctcg gagaatgagg
     4021 atttccgctg cattgctggc atgtgtgtgg atgctcgtgg tgatctcatc gtggctgaca
     4081 gtagtcgcaa ggaaattctc cattttccta agggtggggg ctatagtgtc cttattcgag
     4141 agggacttac ctgtccggtg ggcatagccc taactcctaa ggggcagctg ctggtcttgg
     4201 actgttggga tcattgcatc aagatctaca gctaccatct gagaagatat tccaccccag
     4261 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4321 acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     4381 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     4441 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     4501 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     4561 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     4621 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     4681 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     4741 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     4801 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     4861 acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc
     4921 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     4981 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     5041 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5101 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5161 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5221 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5281 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5341 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5401 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5461 acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa
     5521 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5581 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5641 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5701 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5761 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5821 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5881 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     5941 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6001 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6061 cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc
     6121 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     6181 ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     6241 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6301 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6361 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6421 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6481 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6541 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     6601 ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6661 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6721 tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg
     6781 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6841 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6901 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6961 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7021 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7621 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7861 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7921 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7981 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8041 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8101 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8161 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8221 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8281 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8341 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8401 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8461 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8521 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8581 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8641 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8701 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8761 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8821 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8881 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8941 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     9001 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     9061 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     9121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     9181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     9241 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     9301 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     9361 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
