LOCUS       V039902                 9512 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039902
VERSION     V039902
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             2303..3925
                     /label="SLC1A5(NM_005628)"
                     /note="SLC1A5(NM_005628)"
                     /gene="SLC1A5"
     CDS             3947..4012
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4055..4266
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4279..4547
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4575..5600
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5607..6195
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6269..6502
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6574..6695
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6714..6849
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6887..6903)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6911..6927
                     /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(6935..6965)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6980..7001
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7289..7877)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8048..8908)
                     /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(8909..9013)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9487..9503
                     /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 ggatccgcca ccatggtggc cgatcctcct cgagactcca aggggctcgc
     2341 agcggcggag cccaccgcca acgggggcct ggcgctggcc tccatcgagg accaaggcgc
     2401 ggcagcaggc ggctactgcg gttcccggga ccaggtgcgc cgctgccttc gagccaacct
     2461 gcttgtgctg ctgacagtgg tggccgtggt ggccggcgtg gcgctgggac tgggggtgtc
     2521 gggggccggg ggtgcgctgg cgttgggccc ggagcgcttg agcgccttcg tcttcccggg
     2581 cgagctgctg ctgcgtctgc tgcggatgat catcttgccg ctggtggtgt gcagcttgat
     2641 cggcggcgcc gccagcctgg accccggcgc gctcggccgt ctgggcgcct gggcgctgct
     2701 ctttttcctg gtcaccacgc tgctggcgtc ggcgctcgga gtgggcttgg cgctggctct
     2761 gcagccgggc gccgcctccg ccgccatcaa cgcctccgtg ggagccgcgg gcagtgccga
     2821 aaatgccccc agcaaggagg tgctcgattc gttcctggat cttgcgagaa atatcttccc
     2881 ttccaacctg gtgtcagcag cctttcgctc atactctacc acctatgaag agaggaatat
     2941 caccggaacc agggtgaagg tgcccgtggg gcaggaggtg gaggggatga acatcctggg
     3001 cttggtagtg tttgccatcg tctttggtgt ggcgctgcgg aagctggggc ctgaagggga
     3061 gctgcttatc cgcttcttca actccttcaa tgaggccacc atggttctgg tctcctggat
     3121 catgtggtac gcccctgtgg gcatcatgtt cctggtggct ggcaagatcg tggagatgga
     3181 ggatgtgggt ttactctttg cccgccttgg caagtacatt ctgtgctgcc tgctgggtca
     3241 cgccatccat gggctcctgg tactgcccct catctacttc ctcttcaccc gcaaaaaccc
     3301 ctaccgcttc ctgtggggca tcgtgacgcc gctggccact gcctttggga cctcttccag
     3361 ttccgccacg ctgccgctga tgatgaagtg cgtggaggag aataatggcg tggccaagca
     3421 catcagccgt ttcatcctgc ccatcggcgc caccgtcaac atggacggtg ccgcgctctt
     3481 ccagtgcgtg gccgcagtgt tcattgcaca gctcagccag cagtccttgg acttcgtaaa
     3541 gatcatcacc atcctggtca cggccacagc gtccagcgtg ggggcagcgg gcatccctgc
     3601 tggaggtgtc ctcactctgg ccatcatcct cgaagcagtc aacctcccgg tcgaccatat
     3661 ctccttgatc ctggctgtgg actggctagt cgaccggtcc tgtaccgtcc tcaatgtaga
     3721 aggtgacgct ctgggggcag gactcctcca aaattacgtg gaccgtacgg agtcgagaag
     3781 cacagagcct gagttgatac aagtgaagag tgagctgccc ctggatccgc tgccagtccc
     3841 cactgaggaa ggaaaccccc tcctcaaaca ctatcggggg cccgcagggg atgccacggt
     3901 cgcctctgag aaggaatcag tcatgggtgg aggcggatca ctcgaggact acaaagacca
     3961 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc
     4021 gcaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     4081 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     4141 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4201 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4261 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     4321 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     4381 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4441 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4501 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4561 gcgcctacgt cgacatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     4621 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     4681 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     4741 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     4801 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     4861 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     4921 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     4981 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     5041 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     5101 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     5161 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     5221 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     5281 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     5341 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     5401 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     5461 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     5521 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     5581 gtccgagggc aaaggaatag gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     5641 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5701 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5761 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5821 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5881 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5941 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     6001 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6061 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6121 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     6181 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     6241 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa
     6301 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6361 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6421 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6481 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6541 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6601 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6661 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6721 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6781 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6841 ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc
     6901 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6961 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7021 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7081 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8101 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8161 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8221 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8281 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8341 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8401 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8461 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8521 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8581 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8641 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8701 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8761 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8821 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8881 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8941 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9001 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     9061 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     9121 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     9181 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     9241 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     9301 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     9361 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     9421 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     9481 gacgttgtaa aacgacggcc agtgccaagc tg
//
