LOCUS       V033352                 9592 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033352
VERSION     V033352
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..4422
                     /label="COP1(NM_001001740)"
                     /note="COP1(NM_001001740)"
                     /gene="COP1"
     CDS             4429..4518
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4562..4773
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4786..5054
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5081..5680
                     /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    5687..6275
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6349..6582
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6654..6775
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6794..6929
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6967..6983)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6991..7007
                     /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(7015..7045)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7060..7081
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7369..7957)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8128..8988)
                     /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(8989..9093)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9567..9583
                     /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 catgtctggt agccgccagg ccgggtcggg ctccgctggg
     2341 acaagccccg ggtcctcggc ggcctcctcg gtgacttccg cctcctcgtc tttatcctct
     2401 tccccgtcgc cgccttccgt ggcggtttcg gcggcagcgc tggtgtccgg cggggtggcc
     2461 caggccgccg gctcgggcgg cctcgggggc ccggtgcggc ctgtgttggt ggcgcccgcc
     2521 gtatcgggta gcggcggcgg ggcggtgtcc acgggcctgt cccggcacag ctgcgcggcc
     2581 aggcccagcg ccggcgtagg aggcagcagc tccagcctag gcagcggcag caggaagcga
     2641 cctctcctcg cccccctctg caacgggctc atcaactcct acgaggacaa aagcaacgac
     2701 ttcgtatgcc ccatctgctt tgatatgatt gaagaagcat acatgacaaa atgtggccac
     2761 agcttttgct acaagtgtat tcatcagagt ttggaggaca ataatagatg tcccaagtgt
     2821 aactatgttg tggacaatat tgaccatctg tatcctaatt tcttggtgaa tgaactcatt
     2881 cttaaacaga agcaaagatt tgaggaaaag aggttcaaat tggaccactc aaatggccac
     2941 aggtggcaga tatttcaaga ttggttggga actgaccaag ataaccttga tttggccaat
     3001 gtcaatctta tgttggagtt actagtgcag aagaagaaac aactggaagc agaatcacat
     3061 gcagcccaac tacagattct tatggaattc ctcaaggttg caagaagaaa taagagagag
     3121 gaaatgagtg gcttatactc tcctgtcagt gaggatagca cagtgcctca atttgaagct
     3181 ccttctccat cacacagtag tattattgat tccacagaat acagccaacc tccaggtttc
     3241 agtggcagtt ctcagacaaa gaaacagcct tggtataata gcacgttagc atcaagacga
     3301 aaacgactta ctgctcattt tgaagacttg gagcagtgtt acttttctac aaggatgtct
     3361 cgtatctcag atgacagtcg aactgcaagc cagttggatg aatttcagga atgcttgtcc
     3421 aagtttactc gatataattc agtacgacct ttagccacat tgtcatatgc tagtgatctc
     3481 tataatggtt ccagtatagt ctctagtatt gaatttgacc gggattgtga ctattttgcg
     3541 attgctggag ttacaaagaa gattaaagtc tatgaatatg acactgtcat ccaggatgca
     3601 gtggatattc attaccctga gaatgaaatg acctgcaatt cgaaaatcag ctgtatcagt
     3661 tggagtagtt accataagaa cctgttagct agcagtgatt atgaaggcac tgttatttta
     3721 tgggatggat tcacaggaca gaggtcaaag gtctatcagg agcatgagaa gaggtgttgg
     3781 agtgttgact ttaatttgat ggatcctaaa ctcttggctt caggttctga tgatgcaaaa
     3841 gtgaagctgt ggtctaccaa tctagacaac tcagtggcaa gcattgaggc aaaggctaat
     3901 gtgtgctgtg ttaaattcag cccctcttcc agataccatt tggctttcgg ctgtgcagat
     3961 cactgtgtcc actactatga tcttcgtaac actaaacagc caatcatggt attcaaagga
     4021 caccgtaaag cagtctctta tgcaaagttt gtgagtggtg aggaaattgt ctctgcctca
     4081 acagacagtc agctaaaact gtggaatgta gggaaaccat actgcctacg ttccttcaag
     4141 ggtcatatca atgaaaaaaa ctttgtaggc ctggcttcca atggagatta tatagcttgt
     4201 ggaagtgaaa ataactctct ctacctgtac tataaaggac tttctaagac tttgctaact
     4261 tttaagtttg atacagtcaa aagtgttctc gacaaagacc gaaaagaaga tgatacaaat
     4321 gaatttgtta gtgctgtgtg ctggagggca ctaccagatg gggagtccaa tgtgctgatt
     4381 gctgctaaca gtcagggtac aattaaggtg ctagaattgg taggatccga acaaaaacta
     4441 atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga gcaaaagtta
     4501 atttccgagg aggacttata gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4561 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4621 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4681 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4741 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4801 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4861 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4921 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4981 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     5041 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     5101 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     5161 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     5221 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5281 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5341 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5401 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5461 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5521 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5581 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5641 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5701 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5761 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5821 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5881 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5941 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6001 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6061 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     6121 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6181 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6241 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     6301 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6361 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     6421 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6481 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6541 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6601 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6661 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6721 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6781 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6841 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6901 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     6961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7021 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7561 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7801 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7921 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7981 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8041 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8101 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8161 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8221 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8281 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8341 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8401 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8461 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8521 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8581 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8641 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8701 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8761 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8821 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8881 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8941 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9001 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9061 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9121 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     9181 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     9241 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     9301 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     9361 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9421 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9481 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9541 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
