LOCUS       V027874                 8992 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027874
VERSION     V027874
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             2302..3894
                     /label="PKM(NM_002654)"
                     /note="PKM(NM_002654)"
                     /gene="PKM"
     CDS             3901..3918
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3962..4173
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4186..4454
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4481..5080
                     /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    5087..5675
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5749..5982
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6054..6175
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6194..6329
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6367..6383)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6391..6407
                     /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(6415..6445)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6460..6481
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6769..7357)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7528..8388)
                     /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(8389..8493)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8967..8983
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgtcgaag ccccatagtg aagccgggac tgccttcatt
     2341 cagacccagc agctgcacgc agccatggct gacacattcc tggagcacat gtgccgcctg
     2401 gacattgatt caccacccat cacagcccgg aacactggca tcatctgtac cattggccca
     2461 gcttcccgat cagtggagac gttgaaggag atgattaagt ctggaatgaa tgtggctcgt
     2521 ctgaacttct ctcatggaac tcatgagtac catgcggaga ccatcaagaa tgtgcgcaca
     2581 gccacggaaa gctttgcttc tgaccccatc ctctaccggc ccgttgctgt ggctctagac
     2641 actaaaggac ctgagatccg aactgggctc atcaagggca gcggcactgc agaggtggag
     2701 ctgaagaagg gagccactct caaaatcacg ctggataacg cctacatgga aaagtgtgac
     2761 gagaacatcc tgtggctgga ctacaagaac atctgcaagg tggtggaagt gggcagcaag
     2821 atctacgtgg atgatgggct tatttctctc caggtgaagc agaaaggtgc cgacttcctg
     2881 gtgacggagg tggaaaatgg tggctccttg ggcagcaaga agggtgtgaa ccttcctggg
     2941 gctgctgtgg acttgcctgc tgtgtcggag aaggacatcc aggatctgaa gtttggggtc
     3001 gagcaggatg ttgatatggt gtttgcgtca ttcatccgca aggcatctga tgtccatgaa
     3061 gttaggaagg tcctgggaga gaagggaaag aacatcaaga ttatcagcaa aatcgagaat
     3121 catgaggggg ttcggaggtt tgatgaaatc ctggaggcca gtgatgggat catggtggct
     3181 cgtggtgatc taggcattga gattcctgca gagaaggtct tccttgctca gaagatgatg
     3241 attggacggt gcaaccgagc tgggaagcct gtcatctgtg ctactcagat gctggagagc
     3301 atgatcaaga agccccgccc cactcgggct gaaggcagtg atgtggccaa tgcagtcctg
     3361 gatggagccg actgcatcat gctgtctgga gaaacagcca aaggggacta tcctctggag
     3421 gctgtgcgca tgcagcacct gattgcccgt gaggcagagg ctgccatcta ccacttgcaa
     3481 ttatttgagg aactccgccg cctggcgccc attaccagcg accccacaga agccaccgcc
     3541 gtgggtgccg tggaggcctc cttcaagtgc tgcagtgggg ccataatcgt cctcaccaag
     3601 tctggcaggt ctgctcacca ggtggccaga taccgcccac gtgcccccat cattgctgtg
     3661 acccggaatc cccagacagc tcgtcaggcc cacctgtacc gtggcatctt ccctgtgctg
     3721 tgcaaggacc cagtccagga ggcctgggct gaggacgtgg acctccgggt gaactttgcc
     3781 atgaatgttg gcaaggcccg aggcttcttc aagaagggag atgtggtcat tgtgctgacc
     3841 ggatggcgcc ctggctccgg cttcaccaac accatgcgtg ttgttcctgt gccgggatcc
     3901 catcaccatc accatcacta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     3961 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4021 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4081 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4141 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4201 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4261 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4321 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4381 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4441 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     4501 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     4561 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     4621 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4681 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4741 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4801 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4861 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4921 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4981 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5041 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5101 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5161 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5221 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5281 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5341 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5401 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5461 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     5521 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5581 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5641 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     5701 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5761 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5821 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5881 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5941 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6001 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6181 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6301 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     6361 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6421 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6481 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6541 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6601 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6661 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6721 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6781 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6841 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6901 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6961 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7021 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7081 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7141 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7201 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7261 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7321 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7381 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7441 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7501 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7561 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7621 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7681 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7741 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7801 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7861 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7921 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7981 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8041 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8101 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8161 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8221 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8281 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8341 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8401 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8461 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8521 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8581 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8641 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8701 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     8761 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     8821 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     8881 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     8941 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
