LOCUS       V032284                 9359 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032284
VERSION     V032284
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..4191
                     /label="SLC3A2(NM_002394)"
                     /note="SLC3A2(NM_002394)"
                     /gene="SLC3A2"
     CDS             4198..4224
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4228..4254
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4258..4284
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4329..4540
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4553..4821
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4848..5447
                     /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    5454..6042
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6116..6349
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6421..6542
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6561..6696
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6734..6750)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6758..6774
                     /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(6782..6812)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6827..6848
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7136..7724)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7895..8755)
                     /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(8756..8860)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9334..9350
                     /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 catggagcta cagcctcctg aagcctcgat cgccgtcgtg
     2341 tcgattccgc gccagttgcc tggctcacat tcggaggctg gtgtccaggg tctcagcgcg
     2401 ggggacgact cagagttggg gtctcactgt gttgcccaga ctggtctcga actcttggcc
     2461 tcaggtgatc ctcttccctc agcttcccag aatgccgaga tgatagagac ggggtctgac
     2521 tgtgttaccc aggctggtct tcaactcttg gcctcaagtg atcctcctgc cttagcttcc
     2581 aagaatgctg aggttacagg caccatgagc caggacaccg aggtggatat gaaggaggtg
     2641 gagctgaatg agttagagcc cgagaagcag ccgatgaacg cggcgtctgg ggcggccatg
     2701 tccctggcgg gagccgagaa gaatggtctg gtgaagatca aggtggcgga agacgaggcg
     2761 gaggcggcag ccgcggctaa gttcacgggc ctgtccaagg aggagctgct gaaggtggca
     2821 ggcagccccg gctgggtacg cacccgctgg gcactgctgc tgctcttctg gctcggctgg
     2881 ctcggcatgc ttgctggtgc cgtggtcata atcgtgcgag cgccgcgttg tcgcgagcta
     2941 ccggcgcaga agtggtggca cacgggcgcc ctctaccgca tcggcgacct tcaggccttc
     3001 cagggccacg gcgcgggcaa cctggcgggt ctgaaggggc gtctcgatta cctgagctct
     3061 ctgaaggtga agggccttgt gctgggtcca attcacaaga accagaagga tgatgtcgct
     3121 cagactgact tgctgcagat cgaccccaat tttggctcca aggaagattt tgacagtctc
     3181 ttgcaatcgg ctaaaaaaaa gagcatccgt gtcattctgg accttactcc caactaccgg
     3241 ggtgagaact cgtggttctc cactcaggtt gacactgtgg ccaccaaggt gaaggatgct
     3301 ctggagtttt ggctgcaagc tggcgtggat gggttccagg ttcgggacat agagaatctg
     3361 aaggatgcat cctcattctt ggctgagtgg caaaatatca ccaagggctt cagtgaagac
     3421 aggctcttga ttgcggggac taactcctcc gaccttcagc agatcctgag cctactcgaa
     3481 tccaacaaag acttgctgtt gactagctca tacctgtctg attctggttc tactggggag
     3541 catacaaaat ccctagtcac acagtatttg aatgccactg gcaatcgctg gtgcagctgg
     3601 agtttgtctc aggcaaggct cctgacttcc ttcttgccgg ctcaacttct ccgactctac
     3661 cagctgatgc tcttcaccct gccagggacc cctgttttca gctacgggga tgagattggc
     3721 ctggatgcag ctgcccttcc tggacagcct atggaggctc cagtcatgct gtgggatgag
     3781 tccagcttcc ctgacatccc aggggctgta agtgccaaca tgactgtgaa gggccagagt
     3841 gaagaccctg gctccctcct ttccttgttc cggcggctga gtgaccagcg gagtaaggag
     3901 cgctccctac tgcatgggga cttccacgcg ttctccgctg ggcctggact cttctcctat
     3961 atccgccact gggaccagaa tgagcgtttt ctggtagtgc ttaactttgg ggatgtgggc
     4021 ctctcggctg gactgcaggc ctccgacctg cctgccagcg ccagcctgcc agccaaggct
     4081 gacctcctgc tcagcaccca gccaggccgt gaggagggct cccctcttga gctggaacgc
     4141 ctgaaactgg agcctcacga agggctgctg ctccgcttcc cctacgcggc cggatcctac
     4201 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat
     4261 ccttatgatg tcccagacta cgcataggcg gccgcgaagg atctgcgatc gctccggtgc
     4321 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     4381 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     4441 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     4501 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     4561 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     4621 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     4681 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     4741 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     4801 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagatg accgagtaca
     4861 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
     4921 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
     4981 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5041 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     5101 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     5161 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     5221 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     5281 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     5341 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     5401 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc gacaatcaac
     5461 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5521 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5581 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5641 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5701 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5761 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5821 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5881 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5941 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6001 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca
     6061 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     6121 gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg
     6181 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6241 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6301 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6361 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6421 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6481 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6541 tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc
     6601 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6661 tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa
     6721 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6781 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6841 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7561 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8881 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     8941 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     9001 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     9061 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     9121 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     9181 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     9241 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     9301 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
