LOCUS       V033233                 9517 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033233
VERSION     V033233
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..4371
                     /label="SLC34A2(NM_006424)"
                     /note="SLC34A2(NM_006424)"
                     /gene="SLC34A2"
     CDS             4378..4443
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4487..4698
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4711..4979
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5006..5605
                     /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    5612..6200
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6274..6507
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6579..6700
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6719..6854
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6892..6908)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6916..6932
                     /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(6940..6970)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6985..7006
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7294..7882)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8053..8913)
                     /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(8914..9018)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9492..9508
                     /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 catggctccc tggcctgaat tgggagatgc ccagcccaac
     2341 cccgataagt acctcgaagg ggccgcaggt cagcagccca ctgcccctga taaaagcaaa
     2401 gagaccaaca aaacagataa cactgaggca cctgtaacca agattgaact tctgccgtcc
     2461 tactccacgg ctacactgat agatgagccc actgaggtgg atgacccctg gaacctaccc
     2521 actcttcagg actcggggat caagtggtca gagagagaca ccaaagggaa gattctctgt
     2581 ttcttccaag ggattgggag attgatttta cttctcggat ttctctactt tttcgtgtgc
     2641 tccctggata ttcttagtag cgccttccag ctggttggag gaaaaatggc aggacagttc
     2701 ttcagcaaca gctctattat gtccaaccct ttgttggggc tggtgatcgg ggtgctggtg
     2761 accgtcttgg tgcagagctc cagcacctca acgtccatcg ttgtcagcat ggtgtcctct
     2821 tcattgctca ctgttcgggc tgccatcccc attatcatgg gggccaacat tggaacgtca
     2881 atcaccaaca ctattgttgc gctcatgcag gtgggagatc ggagtgagtt cagaagagct
     2941 tttgcaggag ccactgtcca tgacttcttc aactggctgt ccgtgttggt gctcttgccc
     3001 gtggaggtgg ccacccatta cctcgagatc ataacccagc ttatagtgga gagcttccac
     3061 ttcaagaatg gagaagatgc cccagatctt ctgaaagtca tcactaagcc cttcacaaag
     3121 ctcattgtcc agctggataa aaaagttatc agccaaattg caatgaacga tgaaaaagcg
     3181 aaaaacaaga gtcttgtcaa gatttggtgc aaaactttta ccaacaagac ccagattaac
     3241 gtcactgttc cctcgactgc taactgcacc tccccttccc tctgttggac ggatggcatc
     3301 caaaactgga ccatgaagaa tgtgacctac aaggagaaca tcgccaaatg ccagcatatc
     3361 tttgtgaatt tccacctccc ggatcttgct gtgggcacca tcttgctcat actctccctg
     3421 ctggtcctct gtggttgcct gatcatgatt gtcaagatcc tgggctctgt gctcaagggg
     3481 caggtcgcca ctgtcatcaa gaagaccatc aacactgatt tcccctttcc ctttgcatgg
     3541 ttgactggct acctggccat cctcgtcggg gcaggcatga ccttcatcgt acagagcagc
     3601 tctgtgttca cgtcggcctt gacccccctg attggaatcg gcgtgataac cattgagagg
     3661 gcttatccac tcacgctggg ctccaacatc ggcaccacca ccaccgccat cctggccgcc
     3721 ttagccagcc ctggcaatgc attgaggagt tcactccaga tcgccctgtg ccactttttc
     3781 ttcaacatct ccggcatctt gctgtggtac ccgatcccgt tcactcgcct gcccatccgc
     3841 atggccaagg ggctgggcaa catctctgcc aagtatcgct ggttcgccgt cttctacctg
     3901 atcatcttct tcttcctgat cccgctgacg gtgtttggcc tctcgctggc cggctggcgg
     3961 gtgctggttg gtgtcggggt tcccgtcgtc ttcatcatca tcctggtact gtgcctccga
     4021 ctcctgcagt ctcgctgccc acgcgtcctg ccgaagaaac tccagaactg gaacttcctg
     4081 ccgctgtgga tgcgctcgct gaagccctgg gatgccgtcg tctccaagtt caccggctgc
     4141 ttccagatgc gctgctgctg ctgctgccgc gtgtgctgcc gcgcgtgctg cttgctgtgt
     4201 gactgcccca agtgctgccg ctgcagcaag tgctgcgagg acttggagga ggcgcaggag
     4261 gggcaggatg tccctgtcaa ggctcctgag acctttgata acataaccat tagcagagag
     4321 gctcagggtg aggtccctgc ctcggactca aagaccgaat gcacggcctt gggatccgac
     4381 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     4441 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4501 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4561 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4621 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4681 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     4741 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     4801 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     4861 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     4921 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     4981 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     5041 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5101 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5161 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5221 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5281 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5341 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5401 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5461 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5521 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5581 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     5641 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6001 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6121 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6181 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6241 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6301 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6361 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6421 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6481 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6541 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6601 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6661 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6721 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6781 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6841 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6901 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6961 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7021 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7081 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7201 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9061 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9241 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9361 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9421 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9481 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
