LOCUS       V038585                 8983 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038585
VERSION     V038585
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..3837
                     /label="ALDH1A3(NM_000693)"
                     /note="ALDH1A3(NM_000693)"
                     /gene="ALDH1A3"
     CDS             3844..3909
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3953..4164
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4177..4445
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4472..5071
                     /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    5078..5666
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5740..5973
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6045..6166
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6185..6320
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6358..6374)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6382..6398
                     /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(6406..6436)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6451..6472
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6760..7348)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7519..8379)
                     /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(8380..8484)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8958..8974
                     /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 catggccacc gctaacgggg ccgtggaaaa cgggcagccg
     2341 gacaggaagc cgccggccct gccgcgcccc atccgcaacc tggaggtcaa gttcaccaag
     2401 atatttatca acaatgaatg gcacgaatcc aagagtggga aaaagtttgc tacatgtaac
     2461 ccttcaactc gggagcaaat atgtgaagtg gaagaaggag ataagcccga cgtggacaag
     2521 gctgtggagg ctgcacaggt tgccttccag aggggctcgc catggcgccg gctggatgcc
     2581 ctgagtcgtg ggcggctgct gcaccagctg gctgacctgg tggagaggga ccgcgccacc
     2641 ttggccgccc tggagacgat ggatacaggg aagccatttc ttcatgcttt tttcatcgac
     2701 ctggagggct gtattagaac cctcagatac tttgcagggt gggcagacaa aatccagggc
     2761 aagaccatcc ccacagatga caacgtcgtg tgcttcacca ggcatgagcc cattggtgtc
     2821 tgtggggcca tcactccatg gaacttcccc ctgctgatgc tggtgtggaa gctggcaccc
     2881 gccctctgct gtgggaacac catggtcctg aagcctgcgg agcagacacc tctcaccgcc
     2941 ctttatctcg gctctctgat caaagaggcc gggttccctc caggagtggt gaacattgtg
     3001 ccaggattcg ggcccacagt gggagcagca atttcttctc accctcagat caacaagatc
     3061 gccttcaccg gctccacaga ggttggaaaa ctggttaaag aagctgcgtc ccggagcaat
     3121 ctgaagcggg tgacgctgga gctggggggg aagaacccct gcatcgtgtg tgcggacgct
     3181 gacttggact tggcagtgga gtgtgcccat cagggagtgt tcttcaacca aggccagtgt
     3241 tgcacggcag cctccagggt gttcgtggag gagcaggtct actctgagtt tgtcaggcgg
     3301 agcgtggagt atgccaagaa acggcccgtg ggagacccct tcgatgtcaa aacagaacag
     3361 gggcctcaga ttgatcaaaa gcagttcgac aaaatcttag agctgatcga gagtgggaag
     3421 aaggaagggg ccaagctgga atgcgggggc tcagccatgg aagacaaggg gctcttcatc
     3481 aaacccactg tcttctcaga agtcacagac aacatgcgga ttgccaaaga ggagattttc
     3541 gggccagtgc aaccaatact gaagttcaaa agtatcgaag aagtgataaa aagagcgaat
     3601 agcaccgact atggactcac agcagccgtg ttcacaaaaa atctcgacaa agccctgaag
     3661 ttggcttctg ccttagagtc tggaacggtc tggatcaact gctacaacgc cctctatgca
     3721 caggctccat ttggtggctt taaaatgtca ggaaatggca gagaactagg tgaatacgct
     3781 ttggccgaat acacagaagt gaaaactgtc accatcaaac ttggcgacaa gaaccccgga
     3841 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3901 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3961 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     4021 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     4081 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     4141 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     4201 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     4261 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4321 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4381 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4441 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct
     4501 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     4561 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     4621 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4681 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4741 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4801 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4861 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4921 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4981 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5041 ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat
     5101 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5161 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5221 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5281 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5341 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5401 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5461 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5521 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5581 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5641 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     5701 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     5761 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     5821 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     5881 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     5941 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6001 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6181 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6241 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6301 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7201 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8521 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8581 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8641 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8701 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8761 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8821 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     8881 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     8941 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
