LOCUS       V033789                 9172 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033789
VERSION     V033789
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..4026
                     /label="FUT8(NM_178155)"
                     /note="FUT8(NM_178155)"
                     /gene="FUT8"
     CDS             4033..4098
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4142..4353
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4366..4634
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4661..5260
                     /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    5267..5855
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5929..6162
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6234..6355
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6374..6509
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6547..6563)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6571..6587
                     /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(6595..6625)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6640..6661
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6949..7537)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7708..8568)
                     /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(8569..8673)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9147..9163
                     /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 catgcggcca tggactggtt cctggcgttg gattatgctc
     2341 attctttttg cctgggggac cttgctgttt tatataggtg gtcacttggt acgagataat
     2401 gaccatcctg atcactctag ccgagaactg tccaagattc tggcaaagct tgaacgctta
     2461 aaacaacaga atgaagactt gaggcgaatg gccgaatctc tccggatacc agaaggccct
     2521 attgatcagg ggccagctat aggaagagta cgcgttttag aagagcagct tgttaaggcc
     2581 aaagaacaga ttgaaaatta caagaaacag accagaaatg gtctggggaa ggatcatgaa
     2641 atcctgagga ggaggattga aaatggagct aaagagctct ggtttttcct acagagtgaa
     2701 ttgaagaaat taaagaactt agaaggaaat gaactccaaa gacatgcaga tgaatttctt
     2761 ttggatttag gacatcatga aaggtctata atgacggatc tatactacct cagtcagaca
     2821 gatggagcag gtgattggcg ggaaaaagag gccaaagatc tgacagaact ggttcagcgg
     2881 agaataacat atcttcagaa tcccaaggac tgcagcaaag ccaaaaagct ggtgtgtaat
     2941 atcaacaaag gctgtggcta tggctgtcag ctccatcatg tggtctactg cttcatgatt
     3001 gcatatggca cccagcgaac actcatcttg gaatctcaga attggcgcta tgctactggt
     3061 ggatgggaga ctgtatttag gcctgtaagt gagacatgca cagacagatc tggcatctcc
     3121 actggacact ggtcaggtga agtgaaggac aaaaatgttc aagtggtcga gcttcccatt
     3181 gtagacagtc ttcatccccg tcctccatat ttacccttgg ctgtaccaga agacctcgca
     3241 gatcgacttg tacgagtgca tggtgaccct gcagtgtggt gggtgtctca gtttgtcaaa
     3301 tacttgatcc gcccacagcc ttggctagaa aaagaaatag aagaagccac caagaagctt
     3361 ggcttcaaac atccagttat tggagtccat gtcagacgca cagacaaagt gggaacagaa
     3421 gctgccttcc atcccattga agagtacatg gtgcatgttg aagaacattt tcagcttctt
     3481 gcacgcagaa tgcaagtgga caaaaaaaga gtgtatttgg ccacagatga cccttcttta
     3541 ttaaaggagg caaaaacaaa gtaccccaat tatgaattta ttagtgataa ctctatttcc
     3601 tggtcagctg gactgcacaa tcgatacaca gaaaattcac ttcgtggagt gatcctggat
     3661 atacattttc tctctcaggc agacttccta gtgtgtactt tttcatccca ggtctgtcga
     3721 gttgcttatg aaattatgca aacactacat cctgatgcct ctgcaaactt ccattcttta
     3781 gatgacatct actattttgg gggccagaat gcccacaatc aaattgccat ttatgctcac
     3841 caaccccgaa ctgcagatga aattcccatg gaacctggag atatcattgg tgtggctgga
     3901 aatcattggg atggctattc taaaggtgtc aacaggaaat tgggaaggac gggcctatat
     3961 ccctcctaca aagttcgaga gaagatagaa acggtcaagt accccacata tcctgaggct
     4021 gagaaaggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     4081 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4141 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4201 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4261 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4321 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4381 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4441 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4501 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4561 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4621 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     4681 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     4741 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     4801 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4861 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4921 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4981 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5041 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5101 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5161 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5221 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5281 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5341 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5401 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5461 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5521 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5581 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5641 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     5701 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5761 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5821 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     5881 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5941 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     6001 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6061 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6121 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6181 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6361 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6421 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6481 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     6541 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6601 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6661 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6721 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6781 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6841 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7381 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7741 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7801 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7861 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7921 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7981 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8041 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8101 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8161 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8221 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8281 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8341 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8401 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8461 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8521 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8581 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8641 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8701 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8761 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8821 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8881 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     8941 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9001 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9061 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9121 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
