LOCUS       V026207                 9072 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026207
VERSION     V026207
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             2301..3926
                     /label="SLC1A3(NM_004172)"
                     /note="SLC1A3(NM_004172)"
                     /gene="SLC1A3"
     CDS             3933..3998
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4042..4253
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4266..4534
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4561..5160
                     /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    5167..5755
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5829..6062
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6134..6255
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6274..6409
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6447..6463)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6471..6487
                     /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(6495..6525)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6540..6561
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6849..7437)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7608..8468)
                     /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(8469..8573)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9047..9063
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgactaaaa gcaatggaga agagcccaag atggggggca
     2341 ggatggagag attccagcag ggagtccgta aacgcacact tttggccaag aagaaagtgc
     2401 agaacattac aaaggaggat gttaaaagtt acctgtttcg gaatgctttt gtgctgctca
     2461 cagtcaccgc tgtcattgtg ggtacaatcc ttggatttac cctccgacca tacagaatga
     2521 gctaccggga agtcaagtac ttctcctttc ctggggaact tctgatgagg atgttacaga
     2581 tgctggtctt accacttatc atctccagtc ttgtcacagg aatggcggcg ctagatagta
     2641 aggcatcagg gaagatggga atgcgagctg tagtctatta tatgactacc accatcattg
     2701 ctgtggtgat tggcataatc attgtcatca tcatccatcc tgggaagggc acaaaggaaa
     2761 acatgcacag agaaggcaaa attgtacgag tgacagctgc agatgccttc ctggacttga
     2821 tcaggaacat gttccctcca aatctggtag aagcctgctt taaacagttt aaaaccaact
     2881 atgagaagag aagctttaaa gtgcccatcc aggccaacga aacgcttgtg ggtgctgtga
     2941 taaacaatgt gtctgaggcc atggagactc ttacccgaat cacagaggag ctggtcccag
     3001 ttccaggatc tgtgaatgga gtcaatgccc tgggtctagt tgtcttctcc atgtgcttcg
     3061 gttttgtgat tggaaacatg aaggaacagg ggcaggccct gagagagttc tttgattctc
     3121 ttaacgaagc catcatgaga ctggtagcag taataatgtg gtatgccccc gtgggtattc
     3181 tcttcctgat tgctgggaag attgtggaga tggaagacat gggtgtgatt ggggggcagc
     3241 ttgccatgta caccgtgact gtcattgttg gcttactcat tcacgcagtc atcgtcttgc
     3301 cactcctcta cttcttggta acacggaaaa acccttgggt ttttattgga gggttgctgc
     3361 aagcactcat caccgctctg gggacctctt caagttctgc caccctaccc atcaccttca
     3421 agtgcctgga agagaacaat ggcgtggaca agcgcgtcac cagattcgtg ctccccgtag
     3481 gagccaccat taacatggat gggactgccc tctatgaggc tttggctgcc attttcattg
     3541 ctcaagttaa caactttgaa ctgaacttcg gacaaattat tacaatcagc atcacagcca
     3601 cagctgccag tattggggca gctggaattc ctcaggcggg cctggtcact atggtcattg
     3661 tgctgacatc tgtcggcctg cccactgacg acatcacgct catcatcgcg gtggactggt
     3721 tcctggatcg cctccggacc accaccaacg tactgggaga ctccctggga gctgggattg
     3781 tggagcactt gtcacgacat gaactgaaga acagagatgt tgaaatgggt aactcagtga
     3841 ttgaagagaa tgaaatgaag aaaccatatc aactgattgc acaggacaat gaaactgaga
     3901 aacccatcga cagtgaaacc aagatgggat ccgactacaa agaccatgac ggtgattata
     3961 aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga aggatctgcg
     4021 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     4081 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     4141 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     4201 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     4261 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     4321 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     4381 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     4441 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     4501 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     4561 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     4621 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     4681 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4741 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4801 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4861 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     4921 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4981 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5041 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5101 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5161 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5221 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5281 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5341 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5401 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5461 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5521 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     5581 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5641 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5701 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt
     5761 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5821 ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac
     5881 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5941 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6001 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6061 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     6121 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6181 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6241 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc
     6301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6421 agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7261 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7681 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7741 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7801 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7861 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7921 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7981 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8041 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8101 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8161 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8221 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8281 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8341 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8401 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8461 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8581 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     8641 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     8701 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     8761 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     8821 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     8881 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     8941 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     9001 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     9061 agtgccaagc tg
//
