LOCUS       V044476                 9494 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044476
VERSION     V044476
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..3018
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="photoactivatable GFP variant (Patterson et al.,
                     2002)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="PA-GFP"
     CDS             3067..4419
                     /label="TUBA1B(NM_006082)"
                     /note="TUBA1B(NM_006082)"
                     /gene="TUBA1B"
     promoter        4464..4675
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4688..4956
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4983..5582
                     /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    5589..6177
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6251..6484
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6556..6677
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6696..6831
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6869..6885)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6893..6909
                     /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(6917..6947)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6962..6983
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7271..7859)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8030..8890)
                     /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(8891..8995)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9469..9485
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accttcagct acggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc caccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg atccctcgag gggcccggtg gaggcggatc aggaggaggg
     3061 ggaagcatgc gtgagtgcat ctccatccac gttggccagg ctggtgtcca gattggcaat
     3121 gcctgctggg agctctactg cctggaacac ggcatccagc ccgatggcca gatgccaagt
     3181 gacaagacca ttgggggagg agatgactcc ttcaacacct tcttcagtga gacgggcgct
     3241 ggcaagcacg tgccccgggc tgtgtttgta gacttggaac ccacagtcat tgatgaagtt
     3301 cgcactggca cctaccgcca gctcttccac cctgagcagc tcatcacagg caaggaagat
     3361 gctgccaata actatgcccg agggcactac accattggca aggagatcat tgaccttgtg
     3421 ttggaccgaa ttcgcaagct ggctgaccag tgcaccggtc ttcagggctt cttggttttc
     3481 cacagctttg gtgggggaac tggttctggg ttcacctccc tgctcatgga acgtctctca
     3541 gttgattatg gcaagaagtc caagctggag ttctccattt acccagcacc ccaggtttcc
     3601 acagctgtag ttgagcccta caactccatc ctcaccaccc acaccaccct ggagcactct
     3661 gattgtgcct tcatggtaga caatgaggcc atctatgaca tctgtcgtag aaacctcgat
     3721 atcgagcgcc caacctacac taaccttaac cgccttatta gccagattgt gtcctccatc
     3781 actgcttccc tgagatttga tggagccctg aatgttgacc tgacagaatt ccagaccaac
     3841 ctggtgccct acccccgcat ccacttccct ctggccacat atgcccctgt catctctgct
     3901 gagaaagcct accatgaaca gctttctgta gcagagatca ccaatgcttg ctttgagcca
     3961 gccaaccaga tggtgaaatg tgaccctcgc catggtaaat acatggcttg ctgcctgttg
     4021 taccgtggtg acgtggttcc caaagatgtc aatgctgcca ttgccaccat caaaaccaag
     4081 cgcagcatcc agtttgtgga ttggtgcccc actggcttca aggttggcat caactaccag
     4141 cctcccactg tggtgcctgg tggagacctg gccaaggtac agagagctgt gtgcatgctg
     4201 agcaacacca cagccattgc tgaggcctgg gctcgcctgg accacaagtt tgacctgatg
     4261 tatgccaagc gtgcctttgt tcactggtac gtgggtgagg ggatggagga aggcgagttt
     4321 tcagaggccc gtgaagatat ggctgccctt gagaaggatt atgaggaggt tggtgtggat
     4381 tctgttgaag gagagggtga ggaagaagga gaggaatact aagcggccgc gaaggatctg
     4441 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     4501 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     4561 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     4621 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     4681 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     4741 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     4801 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     4861 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     4921 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     4981 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     5041 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     5101 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5161 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5221 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5281 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5341 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5401 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5461 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5521 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     5581 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     5641 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5701 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5761 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5821 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     5881 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     5941 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     6001 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6061 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6121 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     6181 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6241 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt
     6301 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6361 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6421 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6481 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6541 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6601 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6661 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt
     6721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6781 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt
     6841 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6901 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6961 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7021 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7081 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7141 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7201 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7681 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7801 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7861 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7981 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8161 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8341 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8641 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8881 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8941 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9001 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     9061 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     9121 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     9181 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     9241 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     9301 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     9361 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     9421 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     9481 ccagtgccaa gctg
//
