LOCUS       V025757                 7948 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025757
VERSION     V025757
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..2997
                     /label="Flt3l(NM_013520)"
                     /note="Flt3l(NM_013520)"
                     /gene="Flt3l"
     CDS             3004..3069
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3113..3324
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3337..3605
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3638..4036
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    4043..4631
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             4705..4938
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5010..5131
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5150..5285
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5323..5339)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5347..5363
                     /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(5371..5401)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5416..5437
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5725..6313)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6484..7344)
                     /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(7345..7449)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     7923..7939
                     /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 catgacagtg ctggcgccag cctggagccc aaattcctcc
     2341 ctgttgctgc tgttgctgct gctgagtcct tgcctgcggg ggacacctga ctgttacttc
     2401 agccacagtc ccatctcctc caacttcaaa gtgaagttta gagagttgac tgaccacctg
     2461 cttaaagatt acccagtcac tgtggccgtc aatcttcagg acgagaagca ctgcaaggcc
     2521 ttgtggagcc tcttcctagc ccagcgctgg atagagcaac tgaagactgt ggcagggtct
     2581 aagatgcaaa cgcttctgga ggacgtcaac accgagatac attttgtcac ctcatgtacc
     2641 ttccagcccc taccagaatg tctgcgattc gtccagacca acatctccca cctcctgaag
     2701 gacacctgca cacagctgct tgctctgaag ccctgtatcg ggaaggcctg ccagaatttc
     2761 tctcggtgcc tggaggtgca gtgccagccg gactcctcca ccctgctgcc cccaaggagt
     2821 cccatagccc tagaagccac ggagctccca gagcctcggc ccaggcagct gttgctcctg
     2881 ctgctgctgc tgctgcctct cacactggtg ctgctggcag ccgcctgggg ccttcgctgg
     2941 caaagggcaa gaaggagggg ggagctccac cctggggtgc ccctcccctc ccatcccgga
     3001 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3061 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3121 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3181 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3241 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3301 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     3361 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     3421 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     3481 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     3541 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     3601 agatccaagc tgtgaccggc gcctacgcta ggccaccatg gccaagcctt tgtctcaaga
     3661 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     3721 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     3781 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     3841 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     3901 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     3961 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     4021 tgtgtgggag ggctaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4081 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4141 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4201 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4261 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4321 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4381 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4441 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4501 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4561 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4621 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     4681 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc
     4741 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     4801 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     4861 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     4921 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     4981 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     5041 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5101 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     5161 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5221 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5281 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     5341 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5461 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7501 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7561 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     7621 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7681 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7741 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     7801 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     7861 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7921 ttgtaaaacg acggccagtg ccaagctg
//
