LOCUS       V043124                 9802 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043124
VERSION     V043124
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..4632
                     /label="SEMA3D(NM_001384900)"
                     /note="SEMA3D(NM_001384900)"
                     /gene="SEMA3D"
     CDS             4639..4728
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4772..4983
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4996..5264
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5291..5890
                     /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    5897..6485
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6559..6792
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6864..6985
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7004..7139
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7177..7193)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7201..7217
                     /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(7225..7255)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7270..7291
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7579..8167)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8338..9198)
                     /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(9199..9303)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9777..9793
                     /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 catgaatgct aataaagatg aaagacttaa agccagaagc
     2341 caagattttc acctttttcc tgctttgatg atgctaagca tgaccatgtt gtttcttcca
     2401 gtcactggca ctttgaagca aaatattcca agactcaagc taacctacaa agacttgctg
     2461 ctttcaaata gctgtattcc ctttttgggt tcatcagaag gactggattt tcaaactctt
     2521 ctcttagatg aggaaagagg caggctgctc ttgggagcca aagaccacat ctttctactc
     2581 agtctggttg acttaaacaa aaattttaag aagatttatt ggcctgctgc aaaggaacgg
     2641 gtggaattat gtaaattagc tgggaaagat gccaatacag aatgtgcaaa tttcatcaga
     2701 gtacttcagc cctataacaa aactcacata tatgtgtgtg gaactggagc atttcatcca
     2761 atatgtgggt atattgatct tggagtctac aaggaggata ttatattcaa actagacaca
     2821 cataatttgg agtctggcag actgaaatgt cctttcgatc ctcagcagcc ttttgcttca
     2881 gtaatgacag atgagtacct ctactctgga acagcttctg atttccttgg caaagatact
     2941 gcattcactc gatcccttgg gcctactcat gaccaccact acatcagaac tgacatttca
     3001 gagcactact ggctcaatgg agcaaaattt attggaactt tcttcatacc agacacctac
     3061 aatccagatg atgataaaat atatttcttc tttcgtgaat catctcaaga aggcagtacc
     3121 tccgataaaa ccatcctttc tcgagttgga agagtttgta agaatgatgt aggaggacaa
     3181 cgcagcctga taaacaagtg gacgactttt cttaaggcca gactgatttg ctcaattcct
     3241 ggaagtgatg gggcagatac ttactttgat gagcttcaag atatttattt actccccaca
     3301 agagatgaaa gaaatcctgt agtatatgga gtctttacta caaccagctc catcttcaaa
     3361 ggctctgctg tttgtgtgta tagcatggct gacatcagag cagtttttaa tggtccatat
     3421 gctcataagg aaagtgcaga ccatcgttgg gtgcagtatg atgggagaat tccttatcca
     3481 cggcctggta catgtccaag caaaacctat gacccactga ttaagtccac ccgagatttt
     3541 ccagatgatg tcatcagttt cataaagcgg cactctgtga tgtataagtc cgtataccca
     3601 gttgcaggag gaccaacgtt caagagaatc aatgtggatt acagactgac acagatagtg
     3661 gtggatcatg tcattgcaga agatggccag tacgatgtaa tgtttcttgg aacagacatt
     3721 ggaactgtcc tcaaagttgt cagcatttca aaggaaaagt ggaatatgga agaggtagtg
     3781 ctggaggagt tgcagatatt caagcactca tcaatcatct tgaacatgga attgtctctg
     3841 aagcagcaac aattgtacat tggttcccga gatggattgg ttcagctctc cttgcacaga
     3901 tgcgacactt atgggaaagc ttgcgcagac tgttgtcttg ccagagaccc ctactgtgcc
     3961 tgggatggaa atgcatgctc tcgatatgct cctacttcta aaaggagagc tagacgccaa
     4021 gatgtaaaat atggcgaccc aatcacccag tgctgggaca tcgaagacag cattagtcat
     4081 gaaactgctg atgaaaaggt gatttttggc attgaattta actcaacctt tctggaatgt
     4141 atacctaaat cccaacaagc aactattaaa tggtatatcc agaggtcagg ggatgagcat
     4201 cgagaggagt tgaagcccga tgaaagaatc atcaaaacgg aatatgggct actgattcga
     4261 agtttgcaga agaaggattc tgggatgtat tactgcaaag cccaggagca cactttcatc
     4321 cacaccatag tgaagctgac tttgaatgtc attgagaatg aacagatgga aaatacccag
     4381 agggcagagc atgaggaggg gaaggtcaag gatctattgg ctgagtcacg gttgagatac
     4441 aaagactaca tccaaatcct tagcagccca aacttcagcc tcgaccagta ctgcgaacag
     4501 atgtggcaca gggagaagcg gagacagaga aacaaggggg gcccaaagtg gaagcacatg
     4561 caggaaatga agaagaaacg aaatcgaaga catcacagag acctggatga gctccctaga
     4621 gctgtagcca cgggatccga acaaaaacta atatcagagg aagatttgga acagaaacta
     4681 atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata gcggccgcga
     4741 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     4801 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     4861 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     4921 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     4981 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     5041 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     5101 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     5161 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     5221 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     5281 cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5341 ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5401 cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5461 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5521 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5581 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     5641 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     5701 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     5761 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     5821 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     5881 cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5941 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6001 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6061 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6121 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6181 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6241 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6301 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6361 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6421 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6481 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6541 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     6601 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6661 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6721 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6781 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6841 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6901 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6961 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     7021 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7081 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     7141 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     7201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7981 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     9361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     9421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     9481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     9541 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     9601 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     9661 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     9721 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     9781 aacgacggcc agtgccaagc tg
//
