LOCUS       V025741                 9120 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025741
VERSION     V025741
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..3974
                     /label="Glud1(NM_012570)"
                     /note="Glud1(NM_012570)"
                     /gene="Glud1"
     CDS             3981..4046
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4090..4301
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4314..4582
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4609..5208
                     /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    5215..5803
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5877..6110
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6182..6303
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6322..6457
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6495..6511)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6519..6535
                     /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(6543..6573)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6588..6609
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6897..7485)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7656..8516)
                     /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(8517..8621)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9095..9111
                     /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 atgtaccgcc gtctgggcga agtgctgcta ctgtcccgcg
     2341 ccgggcccgc tgccctgggc tctgcggctg cagactcagc cgcactgctg ggctgggctc
     2401 gcggacaacc ctctgctgtc ccgcaacccg ggctcacgcc ggtcgccagg cgccactaca
     2461 gcgaagcggc caccgaccgc gaagacgacc ccaacttctt caagatggtg gagggcttct
     2521 tcgaccgcgg cgccagcatc gtggaggaca agctggtgga agacctgaag acccgggaga
     2581 acgaggagca gaagcggaac cgagtgcgcg gcatcctgcg gatcatcaag ccttgcaacc
     2641 atgtgttgag cctctccttc cccatccggc gcgacgacgg ctcctgggag gtcatcgaag
     2701 gctaccgggc ccagcacagc cagcaccgca cgccctgcaa gggaggtatc cggtacagca
     2761 ctgacgtgag tgtggatgag gtgaaggcgc tggcgtccct aatgacctac aagtgtgcag
     2821 tggttgatgt gccatttgga ggtgctaaag caggcgttaa gatcaatccc aagaactata
     2881 cagataatga attagaaaag attacacgaa gattcaccat ggagctggca aagaagggtt
     2941 ttattggtcc tggcattgat gtgcctgccc cagacatgag cacgggcgag cgggagatgt
     3001 cctggatcgc tgacacctat gccagcacca tagggcacta tgatatcaat gcacacgcct
     3061 gcgtcactgg taaacccatc agccaaggag gcatccacgg acgcatctcc gctactggcc
     3121 ggggtgtttt tcatgggatt gagaacttca tcaatgaagc ttcctacatg agcatcttag
     3181 ggatgacccc ggggcttggc gataagacgt ttgttgttca gggatttggt aatgtgggcc
     3241 tgcactctat gagatattta catcgttttg gtgctaagtg tgttggtgtt ggagaatctg
     3301 atgggagtat atggaatcca gatggtattg acccaaaaga actggaagat ttcaagttgc
     3361 aacatggatc aattctgggc ttccccaaag ccaaggtcta tgaaggaagc atcttggagg
     3421 ctgactgcga cattttaatc cctgcagcca gcgagaagca gttgaccaaa tccaatgcac
     3481 ccagagtcaa agccaagatc attgctgaag gagccaatgg cccaaccact ccagaggccg
     3541 ataagatttt cctagaaaga aacatcatgg ttattccaga tctctacctg aatgctggag
     3601 gagtgacagt atcttacttt gagtggctaa agaatctaaa tcacgtcagc tatggccgat
     3661 tgaccttcaa atatgaaagg gactcgaact accacttgct catgtccgtt caagagagtt
     3721 tagagagaaa gtttggaaag cacggcggga ctatccctgt ggtccccaca gcagagttcc
     3781 aggacagaat atcgggtgca tctgagaaag acatcgtgca ctctggcttg gcctacacaa
     3841 tggagcgatc tgccaggcaa attatgcgca cagccatgaa gtataacctg ggattggacc
     3901 tgagaacagc tgcctacgtc aatgccattg agaaagtctt caaggtgtat aatgaggctg
     3961 gcgtgacctt cacaggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     4021 tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat cgctccggtg
     4081 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     4141 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     4201 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     4261 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     4321 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     4381 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     4441 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     4501 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     4561 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat gaccgagtac
     4621 aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc
     4681 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag
     4741 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4801 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4861 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4921 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4981 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5041 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5101 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5161 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa
     5221 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5281 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5341 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5401 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5461 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5521 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5581 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5641 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5701 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5761 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc
     5821 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     5881 agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg
     5941 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6001 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6061 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6121 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6181 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6241 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6301 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg
     6361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6421 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca
     6481 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6541 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6601 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6661 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6721 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6781 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6841 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6901 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6961 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7021 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7081 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7141 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7201 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7261 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7321 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7381 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7441 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7501 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7561 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7621 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7681 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7741 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7801 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7861 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7921 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7981 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8041 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8101 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8161 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8221 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8281 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8341 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8401 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8461 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8521 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8581 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8641 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     8701 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     8761 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     8821 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     8881 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     8941 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     9001 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     9061 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
