LOCUS       V031744                 9209 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031744
VERSION     V031744
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..4041
                     /label="SLC5A7(NM_021815)"
                     /note="SLC5A7(NM_021815)"
                     /gene="SLC5A7"
     CDS             4048..4074
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4078..4104
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4108..4134
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4179..4390
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4403..4671
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4698..5297
                     /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    5304..5892
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5966..6199
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6271..6392
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6411..6546
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6584..6600)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6608..6624
                     /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(6632..6662)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6677..6698
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6986..7574)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7745..8605)
                     /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(8606..8710)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9184..9200
                     /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 catggctttc catgtggaag gactgatagc tatcatcgtg
     2341 ttctaccttc taattttgct ggttggaata tgggctgcct ggagaaccaa aaacagtggc
     2401 agcgcagaag agcgcagcga agccatcata gttggtggcc gagatattgg tttattggtt
     2461 ggtggattta ccatgacagc tacctgggtc ggaggagggt atatcaatgg cacagctgaa
     2521 gcagtttatg taccaggtta tggcctagct tgggctcagg caccaattgg atattctctt
     2581 agtctgattt taggtggcct gttctttgca aaacctatgc gttcaaaggg gtatgtgacc
     2641 atgttagacc cgtttcagca aatctatgga aaacgcatgg gcggactcct gtttattcct
     2701 gcactgatgg gagaaatgtt ctgggctgca gcaattttct ctgctttggg agccaccatc
     2761 agcgtgatca tcgatgtgga tatgcacatt tctgtcatca tctctgcact cattgccact
     2821 ctgtacacac tggtgggagg gctctattct gtggcctaca ctgatgtcgt tcagctcttt
     2881 tgcatttttg tagggctgtg gatcagcgtc ccctttgcat tgtcacatcc tgcagtcgca
     2941 gacatcgggt tcactgctgt gcatgccaaa taccaaaagc cgtggctggg aactgttgac
     3001 tcatctgaag tctactcttg gcttgatagt tttctgttgt tgatgctggg tggaatccca
     3061 tggcaagcat actttcagag ggttctctct tcttcctcag ccacctatgc tcaagtgctg
     3121 tccttcctgg cagctttcgg gtgcctggtg atggccatcc cagccatact cattggggcc
     3181 attggagcat caacagactg gaaccagact gcatatgggc ttccagatcc caagactaca
     3241 gaagaggcag acatgatttt accaattgtt ctgcagtatc tctgccctgt gtatatttct
     3301 ttctttggtc ttggtgcagt ttctgctgct gttatgtcat cagcagattc ttccatcttg
     3361 tcagcaagtt ccatgtttgc acggaacatc taccagcttt ccttcagaca aaatgcttcg
     3421 gacaaagaaa tcgtttgggt tatgcgaatc acagtgtttg tgtttggagc atctgcaaca
     3481 gccatggcct tgctgacgaa aactgtgtat gggctctggt acctcagttc tgaccttgtt
     3541 tacatcgtta tcttccccca gctgctttgt gtactctttg ttaagggaac caacacctat
     3601 ggggccgtgg caggttatgt ttctggcctc ttcctgagaa taactggagg ggagccatat
     3661 ctgtatcttc agcccttgat cttctaccct ggctattacc ctgatgataa tggtatatat
     3721 aatcagaaat ttccatttaa aacacttgcc atggttacat cattcttaac caacatttgc
     3781 atctcctatc tagccaagta tctatttgaa agtggaacct tgccacctaa attagatgta
     3841 tttgatgctg ttgttgcaag acacagtgaa gaaaacatgg ataagacaat tcttgtcaaa
     3901 aatgaaaata ttaaattaga tgaacttgca cttgtgaagc cacgacagag catgaccctc
     3961 agctcaactt tcaccaataa agaggccttc cttgatgttg attccagtcc agaagggtct
     4021 gggactgaag ataatttaca gggatcctac ccatacgacg tacctgatta tgcaacttat
     4081 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcataggcg
     4141 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     4201 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg
     4261 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     4321 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     4381 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac
     4441 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc
     4501 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc
     4561 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct
     4621 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg
     4681 accggcgcct acgctagatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     4741 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     4801 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     4861 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     4921 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     4981 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     5041 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     5101 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     5161 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5221 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5281 gcaagcccgg tgcctgagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     5341 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5401 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5461 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5521 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5581 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5641 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     5701 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5761 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5821 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5881 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5941 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat
     6001 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6061 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     6121 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6181 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     6241 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     6301 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6361 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     6421 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6481 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6541 gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg
     6601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6781 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6841 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     7441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7501 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7561 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7621 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7681 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7741 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7861 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8041 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8581 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8701 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     8761 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     8821 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     8881 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     8941 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     9001 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     9061 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     9121 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     9181 gttgtaaaac gacggccagt gccaagctg
//
