LOCUS       V028568                 9001 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028568
VERSION     V028568
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             2302..3855
                     /label="Pfkfb2(NM_008825)"
                     /note="Pfkfb2(NM_008825)"
                     /gene="Pfkfb2"
     CDS             3862..3927
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3971..4182
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4195..4463
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4490..5089
                     /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    5096..5684
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5758..5991
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6063..6184
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6203..6338
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6376..6392)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6400..6416
                     /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(6424..6454)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6469..6490
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6778..7366)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7537..8397)
                     /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(8398..8502)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8976..8992
                     /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 agagctagcg aattcgccac catgtctgag aatagtacat tttccccaga agactgcaac
     2341 agcagctata aaccccacgc ctcaaatctg cgaagggcag ggaagacatg ctcatgggct
     2401 tcctatatga ccaactcccc aacactcatt gttatgattg gcttgccagc ccggggtaaa
     2461 acttatgtgt ctaagaaact aacacgctac ctcaactgga ttggagtacc caccaaagtg
     2521 tttaatcttg gggtatatcg acgtgaagca gtcaagtcct atcagtctta tgatttcttt
     2581 cgacatgaca atgaagaggc tatgaagatc cgcaaacagt gtgctctggt ggcactggaa
     2641 gatgttaaag cctattttac tgaagagagt gggcagattg cggtgtttga tgccaccaat
     2701 accactcggg agaggaggga catgattttg aactttgcca agcagaatgc cttcaaggta
     2761 ttttttgtag aatctgtctg tgatgatcct gatgtcattg ctgccaatat tctggaggta
     2821 aaagtgtcga gccctgacta ccccgaaagg aatagggaga atgtgatgga ggacttcctg
     2881 aagagaattg agtgctacaa ggtcacttac cagccccttg acccagacaa ctatgataag
     2941 gatctttctt tcataaaggt gataaatgta ggccagagat tcctggtcaa cagagttcag
     3001 gactacatcc agagtaagat tgtctactac ctgatgaata tccacgtcca tcctcgcacc
     3061 atctaccttt gccggcacgg cgagagcgag ttcaaccttt tggggaagat tgggggtgac
     3121 tctggcctct ccgtgcgagg aaagcagttt gctcatgctc tgaagaagtt tctggaggaa
     3181 caggagatcc aggaccttaa agtgtggacg agccagttga agaggacaat tcagactgct
     3241 gagtctcttg gggtgaccta tgagcagtgg aagatcctca atgagattga tgctggcgtg
     3301 tgtgaggaga tgacatattc agagattgag caacggtacc cagaggaatt tgcactccga
     3361 gatcaagaga agtatctgta ccgataccct ggtggggagt cctaccagga cctggtgcag
     3421 cggctggagc ctgtcatcat ggagctagag cgtcaaggca acatcctcgt tatctctcac
     3481 caggctgtca tgcgctgcct cctggcctac ttcttggata aaggcgcaga tgagttacca
     3541 tacttgaggt gccccctgca catcatcttc aaacttactc ctgtggccta tggttgcaaa
     3601 gtggaaacaa ttactctgaa tgtggatgct gtagacacac atcgtgacaa gccaactcac
     3661 aacttcccca agagccaaac ccctgtaagg atgagaagga acagctttac gcctctgtcc
     3721 agttcgaaca caataaggcg tccaagaaat tacagtgttg ggagccgacc cctcaagccc
     3781 ctcagccctc tccgtgctct ggacatgcaa gaaggggcgg accagccgaa gacccaagtc
     3841 agcattccgg tggtgggatc cgactacaaa gaccatgacg gtgattataa agatcatgac
     3901 atcgactaca aggatgacga tgacaagtag cggccgcgaa ggatctgcga tcgctccggt
     3961 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     4021 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     4081 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     4141 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     4201 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     4261 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     4321 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4381 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4441 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta
     4501 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     4561 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     4621 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4681 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4741 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4801 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4861 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4921 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4981 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5041 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca
     5101 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5161 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5221 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5281 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5341 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5401 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5461 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     5521 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5581 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5641 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     5701 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5761 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     5821 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5881 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5941 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6001 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6061 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6121 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6181 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     6241 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6301 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6361 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6421 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6481 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6541 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6601 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6661 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6721 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6781 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6841 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6901 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6961 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7021 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7081 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7141 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7201 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7261 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7321 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7381 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7441 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7501 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7561 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7621 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7681 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7741 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7801 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7861 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7921 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7981 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8041 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8101 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8161 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8221 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8281 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8341 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8401 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8461 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8521 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8581 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     8641 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     8701 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     8761 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     8821 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     8881 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     8941 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9001 g
//
