LOCUS       V032597                 8752 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032597
VERSION     V032597
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..3654
                     /label="TUBG1(NM_001070)"
                     /note="TUBG1(NM_001070)"
                     /gene="TUBG1"
     CDS             3661..3678
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3722..3933
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3946..4214
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4241..4840
                     /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    4847..5435
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5509..5742
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5814..5935
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5954..6089
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6127..6143)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6151..6167
                     /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(6175..6205)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6220..6241
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6529..7117)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7288..8148)
                     /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(8149..8253)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8727..8743
                     /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 catgccgagg gaaatcatca ccctacagtt gggccagtgc
     2341 ggcaatcaga ttgggttcga gttctggaaa cagctgtgcg ccgagcatgg tatcagcccc
     2401 gagggcatcg tggaggagtt cgccaccgag ggcactgacc gcaaggacgt ctttttctac
     2461 caggcagacg atgagcacta catcccccgg gccgtgctgc tggacttgga accccgggtg
     2521 atccactcca tcctcaactc cccctatgcc aagctctaca acccagagaa catctacctg
     2581 tcggaacatg gaggaggagc tggcaacaac tgggccagcg gattctccca gggagaaaag
     2641 atccatgagg acatttttga catcatagac cgggaggcag atggtagtga cagtctagag
     2701 ggctttgtgc tgtgtcactc cattgctggg gggacaggct ctggactggg ttcctacctc
     2761 ttagaacggc tgaatgacag gtatcctaag aagctggtgc agacatactc agtgtttccc
     2821 aaccaggacg agatgagcga tgtggtggtc cagccttaca attcactcct cacactcaag
     2881 aggctgacgc agaatgcaga ctgtgtggtg gtgctggaca acacagccct gaaccggatt
     2941 gccacagacc gcctgcacat ccagaaccca tccttctccc agatcaacca gctggtgtct
     3001 accatcatgt cagccagcac caccaccctg cgctaccctg gctacatgaa caatgacctc
     3061 atcggcctca tcgcctcgct cattcccacc ccacggctcc acttcctcat gaccggctac
     3121 acccctctca ctacggacca gtcagtggcc agcgtgagga agaccacggt cctggatgtc
     3181 atgaggcggc tgctgcagcc caagaacgtg atggtgtcca caggccgaga ccgccagacc
     3241 aaccactgct acatcgccat cctcaacatc atccagggag aggtggaccc cacccaggtc
     3301 cacaagagct tgcagaggat ccgggaacgc aagttggcca acttcatccc gtggggcccc
     3361 gccagcatcc aggtggccct gtcgaggaag tctccctacc tgccctcggc ccaccgggtc
     3421 agcgggctca tgatggccaa ccacaccagc atctcctcgc tcttcgagag aacctgtcgc
     3481 cagtatgaca agctgcgtaa gcgggaggcc ttcctggagc agttccgcaa ggaggacatg
     3541 ttcaaggaca actttgatga gatggacaca tccagggaga ttgtgcagca gctcatcgat
     3601 gagtaccatg cggccacacg gccagactac atctcctggg gcacccagga gcagggatcc
     3661 catcaccatc accatcacta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     3721 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3781 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3841 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3901 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3961 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4021 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4081 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4141 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4201 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     4261 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     4321 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     4381 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4441 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4501 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4561 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4621 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4681 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4741 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4801 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     4861 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4921 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4981 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5041 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5101 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5161 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5221 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     5281 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5341 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5401 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     5461 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5521 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5581 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5641 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5701 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     5761 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     5821 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5881 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5941 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6061 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     6121 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6181 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6241 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6301 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6361 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6421 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6481 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6961 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7021 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7141 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7201 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7261 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7321 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7381 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7441 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7501 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7561 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7621 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7681 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7741 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7801 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7861 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7921 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7981 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8041 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8101 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8161 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8221 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8281 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8341 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8401 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8461 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     8521 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     8581 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     8641 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     8701 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
