LOCUS       V041597                 9592 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041597
VERSION     V041597
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..4488
                     /label="DCLK1(NM_004734)"
                     /note="DCLK1(NM_004734)"
                     /gene="DCLK1"
     CDS             4495..4518
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     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 catgtccttc ggcagagaca tggagctgga gcacttcgac
     2341 gagcgggata aggcgcagag atacagccga gggtcgcggg tgaacggcct gccgagcccg
     2401 acgcacagcg cccactgcag cttctaccgc acccgcacgc tgcagacgct cagctccgag
     2461 aagaaggcca agaaagttcg tttctatcga aacggagatc gatacttcaa agggattgtg
     2521 tatgccatct ccccagaccg gttccgatct tttgaggccc tgctggctga tttgacccga
     2581 actctgtcgg ataacgtgaa tttgccccag ggagtgagaa caatctacac cattgatggg
     2641 ctcaagaaga tttccagcct ggaccaactg gtggaaggag agagttatgt atgtggctcc
     2701 atagagccct tcaagaaact ggagtacacc aagaatgtga accccaactg gtcggtgaac
     2761 gtcaagacca cctcggcttc tcgggcagtg tcttcactgg ccactgccaa aggaagccct
     2821 tcagaggtgc gagagaataa ggatttcatt cggcccaagc tggtcaccat catcagaagt
     2881 ggcgtgaagc cacggaaagc tgtcaggatt ctgctgaaca agaaaacggc tcattccttt
     2941 gagcaggtcc tcaccgatat caccgatgcc atcaagctgg actcgggagt ggtgaaacgc
     3001 ctgtacacgt tggatgggaa acaggtgatg tgccttcagg acttttttgg tgatgatgac
     3061 atttttattg catgtggacc ggagaagttc cgttaccagg atgatttctt gctagatgaa
     3121 agtgaatgtc gagtggtaaa gtccacttct tacaccaaaa tagcttcatc atcccgcagg
     3181 agcaccacca agagcccagg accgtccagg cgtagcaagt cccctgcctc caccagctca
     3241 gttaatggaa cccctggtag tcagctctct actccgcgct caggcaagtc gccaagccca
     3301 tcacccacca gcccaggaag cctgcggaag cagaggagct ctcagcatgg cggctcctct
     3361 acgtcacttg cgtccaccaa agtctgcagc tcgatggatg agaacgatgg ccctggagaa
     3421 gaagtgtcgg aggaaggctt ccagattcca gctacaataa cagaacgata taaagtcgga
     3481 agaacaatag gagatggaaa ttttgctgtt gtcaaggaat gtgtagaaag atcgactgct
     3541 agagagtatg ctctgaaaat tatcaagaaa agcaaatgtc gaggcaaaga gcacatgatc
     3601 cagaatgaag tgtctatttt aagaagagtg aagcatccca atatcgttct tctgattgag
     3661 gagatggatg tgccaactga actgtatctt gtcatggaat tagtaaaggg gggagacctt
     3721 tttgatgcca ttacttccac taacaaatac accgagagag acgccagtgg gatgctgtac
     3781 aacctagcca gcgccatcaa atacctgcat agcctgaaca tcgtccaccg tgatatcaag
     3841 ccagagaacc tgctggtgta tgagcaccaa gatggcagca aatcactgaa gctgggtgac
     3901 tttggactgg ccaccattgt agacggcccc ctgtacacag tctgtggcac cccaacatac
     3961 gtggctccag aaatcattgc agagactgga tacggcctca aggtggacat ctgggcagca
     4021 ggtgtaatca cttatatcct gctgtgtggt ttccctccat tccgtggaag tggtgatgac
     4081 caggaggtgc tttttgatca gattttgatg gggcaggtgg actttccttc tccatactgg
     4141 gataatgttt ccgattctgc aaaggagctc attaccatga tgctgttggt cgatgtagat
     4201 cagcgatttt ctgctgttca agtacttgag catccctggg ttaatgatga tggcctccca
     4261 gaaaatgaac atcagctgtc agtagctgga aagataaaga agcatttcaa cacaggcccc
     4321 aagccgaata gcacagcagc tggagtttct gtcatagcac tggaccacgg gtttaccatc
     4381 aagagatcag ggtctttgga ctactaccag caaccaggaa tgtattggat aagaccaccg
     4441 ctcttgataa ggagaggcag gttttccgac gaagacgcaa ccaggatgct cgagcaccat
     4501 caccatcacc atcaccatta 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
//
