LOCUS       V018364                 8932 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018364
VERSION     V018364
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             2298..3785
                     /label="SLC2A3(NM_006931)"
                     /note="SLC2A3(NM_006931)"
                     /gene="SLC2A3"
     CDS             3786..3851
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3902..4113
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4126..4394
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4421..5020
                     /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    5027..5615
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5689..5922
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5994..6115
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6134..6269
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6307..6323)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6331..6347
                     /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(6355..6385)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6400..6421
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6709..7297)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7468..8328)
                     /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(8329..8433)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8907..8923
                     /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 agagctagca ggccaccatg gggacacaga aggtcacccc agctctgata tttgccatca
     2341 cagttgctac aatcggctct ttccaatttg gctacaacac tggggtcatc aatgctcctg
     2401 agaagatcat aaaggaattt atcaataaaa ctttgacgga caagggaaat gccccaccct
     2461 ctgaggtgct gctcacgtct ctctggtcct tgtctgtggc catattttcc gtcgggggta
     2521 tgatcggctc cttttccgtc ggactcttcg tcaaccgctt tggcaggcgc aattcaatgc
     2581 tgattgtcaa cctgttggct gtcactggtg gctgctttat gggactgtgt aaagtagcta
     2641 agtcggttga aatgctgatc ctgggtcgct tggttattgg cctcttctgc ggactctgca
     2701 caggttttgt gcccatgtac attggagaga tctcgcctac tgccctgcgg ggtgcctttg
     2761 gcactctcaa ccagctgggc atcgttgttg gaattctggt ggcccagatc tttggtctgg
     2821 aattcatcct tgggtctgaa gagctatggc cgctgctact gggttttacc atccttcctg
     2881 ctatcctaca aagtgcagcc cttccatttt gccctgaaag tcccagattt ttgctcatta
     2941 acagaaaaga agaggagaat gctaagcaga tcctccagcg gttgtggggc acccaggatg
     3001 tatcccaaga catccaggag atgaaagatg agagtgcaag gatgtcacaa gaaaagcaag
     3061 tcaccgtgct agagctcttt agagtgtcca gctaccgaca gcccatcatc atttccattg
     3121 tgctccagct ctctcagcag ctctctggga tcaatgctgt gttctattac tcaacaggaa
     3181 tcttcaagga tgcaggtgtt caagagccca tctatgccac catcggcgcg ggtgtggtta
     3241 atactatctt cactgtagtt tctctatttc tggtggaaag ggcaggaaga aggactctgc
     3301 atatgatagg ccttggaggg atggcttttt gttccacgct catgactgtt tctttgttat
     3361 taaaggataa ctataatggg atgagctttg tctgtattgg ggctatcttg gtctttgtag
     3421 ccttctttga aattggacca ggccccattc cctggtttat tgtggccgaa ctcttcagcc
     3481 agggcccccg cccagctgcg atggcagtgg ccggctgctc caactggacc tccaacttcc
     3541 tagtcggatt gctcttcccc tccgctgctc actatttagg agcctacgtt tttattatct
     3601 tcaccggctt cctcattacc ttcttggctt ttaccttctt caaagtccct gagacccgtg
     3661 gcaggacttt tgaggatatc acacgggcct ttgaagggca ggcacacggt gcagatagat
     3721 ctggaaagga cggcgtcatg gagatgaaca gcatcgagcc tgctaaggag accaccacca
     3781 atgtcgacta caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg
     3841 acgatgacaa gtaaggatcc gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     3901 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3961 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4021 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4081 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4141 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4201 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4261 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4321 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4381 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     4441 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     4501 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     4561 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4621 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4681 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4741 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4801 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4861 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4921 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4981 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5041 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5101 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5161 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5221 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5281 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5341 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5401 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     5461 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5521 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5581 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     5641 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5701 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5761 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5821 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5881 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     5941 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6121 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6181 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6241 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     6301 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6361 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6421 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6481 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6541 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6601 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6661 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6721 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6781 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6841 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6901 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6961 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7021 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7081 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7141 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7201 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7261 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7321 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7381 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7441 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7501 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7561 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7621 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7681 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7741 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7801 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7861 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7921 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7981 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8041 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8101 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8161 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8221 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8281 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8341 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8461 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8521 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8581 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8641 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     8701 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     8761 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     8821 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     8881 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
