LOCUS       V032463                10261 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032463
VERSION     V032463
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..5115
                     /label="Cblb(NM_001033238)"
                     /note="Cblb(NM_001033238)"
                     /gene="Cblb"
     CDS             5122..5187
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5231..5442
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5455..5723
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5750..6349
                     /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    6356..6944
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7018..7251
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7323..7444
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7463..7598
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7636..7652)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7660..7676
                     /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(7684..7714)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7729..7750
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8038..8626)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8797..9657)
                     /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(9658..9762)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10236..10252
                     /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 catggcaaat tctatgaatg gcagaaatcc tggtggtcga
     2341 ggaggaaacc cccgcaaagg tcgcattttg gggattattg atgctattca ggatgcagtc
     2401 ggacccccaa agcaagctgc agctgaccgc aggactgtgg agaagacttg gaaactcatg
     2461 gacaaagtgg taagactgtg ccaaaatccc aagcttcagt tgaaaaacag cccaccgtat
     2521 atacttgata ttttacctga tacgtatcag cacttgagac ttatattgag taaatatgat
     2581 gacaaccaga agctggctca actgagcgag aatgagtact ttaaaatcta catcgatagt
     2641 ctcatgaaga agtcgaagcg agcgatccgg ctctttaaag aaggcaagga aaggatgtac
     2701 gaagagcagt cgcaggacag acggaatctc acaaagctgt cccttatctt cagtcacatg
     2761 ctggcagaaa tcaaggcgat ctttcccaat ggccagttcc agggagataa cttccggatc
     2821 accaaagcag atgctgctga gttctggagg aagttttttg gagacaaaac tattgtacca
     2881 tggaaagtct tcagacagtg cctgcatgag gtccatcaga tcagctctgg cctggaagca
     2941 atggctctga agtcaaccat tgatttaact tgcaatgatt acatctcagt gtttgaattt
     3001 gatattttta ccaggctatt tcagccctgg ggctctattt tacggaattg gaacttcttg
     3061 gctgtgactc acccgggata catggcattt ctcacgtatg atgaagttaa agctcggcta
     3121 cagaaataca gcaccaagcc tggaagttac attttccggt taagctgcac tcggctggga
     3181 caatgggcca ttggctatgt gactggggac ggcaatatcc tacagaccat acctcataac
     3241 aagcccctgt tccaagccct gattgatggt agcagggaag gcttttatct ttatcctgat
     3301 gggcggagct ataaccctga tttaaccgga ttatgtgaac ctacacctca cgatcatata
     3361 aaagttacac aggagcagta tgaactgtat tgtgaaatgg gctccacttt tcagctctgc
     3421 aagatctgtg cagagaatga caaagatgtc aagattgagc cttgcgggca tctgatgtgc
     3481 acttcgtgcc ttaccgcgtg gcaggagtct gatggccaag gctgcccctt ctgtcgctgt
     3541 gagataaaag gaacggagcc catcattgtg gaccccttcg accccagaga tgaaggctcc
     3601 aggtgctgca gcatcattga ccctttcagc atccccatgc ttgacttgga cgatgacgat
     3661 gatcgagagg agtctttgat gatgaacagg ctggcgagtg ttcgaaagtg cacggacagg
     3721 cagaactcac cagtcacatc gccaggctcc tcaccccttg cccagagaag aaagcctcag
     3781 ccagaccctc tccagatccc ccacctcagc ctgccaccag tgcctccccg cctagatctc
     3841 attcagaaag gcatcgtgcg ctctccgtgt ggcagcccca caggctcgcc aaagtcttct
     3901 ccatgcatgg ttagaaaaca agacaaacca ctcccggcac cccctcctcc cttgagagat
     3961 ccacctcctc caccagagag gcctcccccc atcccacctg acaatagact gagcagacac
     4021 ttccaccatg gagagagtgt gccttcccgg gaccagccca tgcccctcga agcctggtgc
     4081 cctcgggatg ccttcgggac taaccaggtg atgggatgtc gcatcctcgg ggatggctct
     4141 ccaaagcctg gcgtcaccgc aaactccagc ttaaatggga ggcacagtag aatgggctcc
     4201 gagcaggttc ttatgaggaa acacagacgc catgatttgc cttcagaagg agccaaggtc
     4261 ttttccaatg gccaccttgc cactgaagaa tacgatgtcc ctcctcggct ttcccctccg
     4321 cctccagtca ccacccttct cccaagcata aagtgtactg gtccattagc aaattgtctc
     4381 tcagagaaaa ctagagacac ggtagaagat gatgacgatg aatacaagat tccttcatcc
     4441 catcctgtgt ccctgaattc acaaccatct cattgccata atgtcaaagc tcctgttcgg
     4501 tcttgtgata atggtcattg tatattgaat ggaactcatg gtgcgccttc agagatgaag
     4561 aaatcgaaca tcccagattt aggcatctat ttgaagggtg aggacgcctt tgatgctctc
     4621 cctccatccc tccctcctcc cccacctcct gcaagacaca gtctcatcga acattcaaaa
     4681 cctccaggct ccagtagccg gccttcctca gggcaggacc ttttccttct tccttcagat
     4741 cccttttttg acccaacaag tggccaagtt ccattgcctc ctgccaggag agcagcagga
     4801 gacagcggca aagccaacag agcctcgcag gactatgacc agctcccttc atcttccgat
     4861 ggttcgcaag caccagctag accccccaaa ccacgacccc gaaggactgc accagaaatt
     4921 catcacagaa agccccatgg gcccgaagcg gcgttggaaa atgtcgatgc aaaaattgca
     4981 aaactcatgg gagaggggta tgcctttgaa gaggtgaaga gagccttaga gattgcccag
     5041 aataacgtgg aagtggccag gagcatactt cgagaatttg ccttcccccc tcctgtctcc
     5101 ccacgtctga atctaggatc cgactacaaa gaccatgacg gtgattataa agatcatgac
     5161 atcgactaca aggatgacga tgacaagtag cggccgcgaa ggatctgcga tcgctccggt
     5221 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     5281 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     5341 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     5401 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     5461 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     5521 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     5581 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     5641 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     5701 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta
     5761 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     5821 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     5881 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5941 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     6001 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     6061 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     6121 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     6181 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     6241 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     6301 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca
     6361 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6421 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6481 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6541 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6601 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6661 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6721 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6781 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6841 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6901 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6961 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     7021 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     7081 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7141 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7201 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7261 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7321 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7381 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7441 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     7501 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     7561 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     7621 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7681 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7741 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7801 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7861 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7921 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7981 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8041 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8101 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8161 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8221 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8281 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8341 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8401 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8461 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8521 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8581 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8641 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8701 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8761 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8821 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8881 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8941 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9001 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9061 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9781 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9841 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9901 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9961 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
    10021 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
    10081 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
    10141 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
    10201 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
    10261 g
//
