LOCUS       V032874                 9513 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032874
VERSION     V032874
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"
     regulatory      2288..2297
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2297..2362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2369..4438
                     /label="ADGRL4(NM_022159)"
                     /note="ADGRL4(NM_022159)"
                     /gene="ADGRL4"
     promoter        4483..4694
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4707..4975
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5002..5601
                     /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    5608..6196
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6270..6503
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6575..6696
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6715..6850
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6888..6904)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6912..6928
                     /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(6936..6966)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6981..7002
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7290..7878)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8049..8909)
                     /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(8910..9014)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9488..9504
                     /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca aggaattcat gaaacgcctc ccgctcctag tggttttttc
     2401 cactttgttg aattgttcct atactcaaaa ttgcaccaag acaccttgtc tcccaaatgc
     2461 aaaatgtgaa atacgcaatg gaattgaagc ctgctattgc aacatgggat tttcaggaaa
     2521 tggtgtcaca atttgtgaag atgataatga atgtggaaat ttaactcagt cctgtggcga
     2581 aaatgctaat tgcactaaca cagaaggaag ttattattgt atgtgtgtac ctggcttcag
     2641 atccagcagt aaccaagaca ggtttatcac taatgatgga accgtctgta tagaaaatgt
     2701 gaatgcaaac tgccatttag ataatgtctg tatagctgca aatattaata aaactttaac
     2761 aaaaatcaga tccataaaag aacctgtggc tttgctacaa gaagtctata gaaattctgt
     2821 gacagatctt tcaccaacag atataattac atatatagaa atattagctg aatcatcttc
     2881 attactaggt tacaagaaca acactatctc agccaaggac accctttcta actcaactct
     2941 tactgaattt gtaaaaaccg tgaataattt tgttcaaagg gatacatttg tagtttggga
     3001 caagttatct gtgaatcata ggagaacaca tcttacaaaa ctcatgcaca ctgttgaaca
     3061 agctacttta aggatatccc agagcttcca aaagaccaca gagtttgata caaattcaac
     3121 ggatatagct ctcaaagttt tcttttttga ttcatataac atgaaacata ttcatcctca
     3181 tatgaatatg gatggagact acataaatat atttccaaag agaaaagctg catatgattc
     3241 aaatggcaat gttgcagttg catttgtata ttataagagt attggtcctt tgctttcatc
     3301 atctgacaac ttcttattga aacctcaaaa ttatgataat tctgaagagg aggaaagagt
     3361 catatcttca gtaatttcag tctcaatgag ctcaaaccca cccacattat atgaacttga
     3421 aaaaataaca tttacattaa gtcatcgaaa ggtcacagat aggtatagga gtctatgtgc
     3481 attttggaat tactcacctg ataccatgaa tggcagctgg tcttcagagg gctgtgagct
     3541 gacatactca aatgagaccc acacctcatg ccgctgtaat cacctgacac attttgcaat
     3601 tttgatgtcc tctggtcctt ccattggtat taaagattat aatattctta caaggatcac
     3661 tcaactagga ataattattt cactgatttg tcttgccata tgcattttta ccttctggtt
     3721 cttcagtgaa attcaaagca ccaggacaac aattcacaaa aatctttgct gtagcctatt
     3781 tcttgctgaa cttgtttttc ttgttgggat caatacaaat actaataagc tcttctgttc
     3841 aatcattgcc ggactgctac actacttctt tttagctgct tttgcatgga tgtgcattga
     3901 aggcatacat ctctatctca ttgttgtggg tgtcatctac aacaagggat ttttgcacaa
     3961 gaatttttat atctttggct atctaagccc agccgtggta gttggatttt cggcagcact
     4021 aggatacaga tattatggca caaccaaagt atgttggctt agcaccgaaa acaactttat
     4081 ttggagtttt ataggaccag catgcctaat cattcttgtt aatctcttgg cttttggagt
     4141 catcatatac aaagtttttc gtcacactgc agggttgaaa ccagaagtta gttgctttga
     4201 gaacataagg tcttgtgcaa gaggagccct cgctcttctg ttccttctcg gcaccacctg
     4261 gatctttggg gttctccatg ttgtgcacgc atcagtggtt acagcttacc tcttcacagt
     4321 cagcaatgct ttccagggga tgttcatttt tttattcctg tgtgttttat ctagaaagat
     4381 tcaagaagaa tattacagat tgttcaaaaa tgtcccctgt tgttttggat gtttaaggta
     4441 agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc
     4501 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt
     4561 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     4621 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     4681 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc
     4741 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg
     4801 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt
     4861 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc
     4921 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc
     4981 tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct cgccacccgc
     5041 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5101 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5161 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5221 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5281 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5341 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5401 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5461 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5521 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5581 acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     5641 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5701 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5761 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5821 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5881 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5941 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6001 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6061 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6121 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6181 tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct
     6241 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata
     6301 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6361 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6421 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6481 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6541 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     6601 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6661 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     6721 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6781 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6841 tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt
     6901 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6961 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7021 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7081 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8161 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8281 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8341 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8401 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8461 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8521 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8581 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8641 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8701 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8761 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8821 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8881 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9001 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     9061 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     9121 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     9181 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     9241 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     9301 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     9361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     9421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     9481 cgacgttgta aaacgacggc cagtgccaag ctg
//
