LOCUS       V025532                 9420 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025532
VERSION     V025532
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..4274
                     /label="KLHL13(NM_001168299)"
                     /note="KLHL13(NM_001168299)"
                     /gene="KLHL13"
     CDS             4281..4346
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4390..4601
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4614..4882
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4909..5508
                     /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    5515..6103
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6177..6410
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6482..6603
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6622..6757
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6795..6811)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6819..6835
                     /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(6843..6873)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6888..6909
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7197..7785)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7956..8816)
                     /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(8817..8921)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9395..9411
                     /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 atgatgagag ttcaaacctt aagagaaaaa tgggcttatt
     2341 ggagacggag gcaattgtca ttaaaacaag ccgatttcaa ggatatattc aaaaaatcaa
     2401 cttcaggatc tctcgtggaa gaggaagacc aacacatgaa attgtccctt ggaggcagcg
     2461 aaatgggcct ctcatcccat ttgcagtctt ccaaggcagg acctacacgc atctttacca
     2521 gcaataccca cagttctgtg gtgttacagg gctttgacca gcttcgactt gaaggattgc
     2581 tttgtgatgt gaccctgatg ccaggtgaca cagatgatgc tttccctgtg catagagtca
     2641 tgatggcatc tgctagtgat tacttcaagg ctatgttcac aggtggaatg aaagaacaag
     2701 atttaatgtg cattaaactt catggtgtga gcaaagtcgg tctaaggaaa attattgatt
     2761 tcatttatac tgcaaagctt tctcttaata tggacaacct tcaagacacg ctggaagctg
     2821 ccagtttcct acagattctg ccagttttgg acttctgtaa agtgtttctc atatctgggg
     2881 tcactttaga caactgtgtt gaagttggac ggattgccaa cacctacaat ctaaccgaag
     2941 tggataaata cgttaacagt ttcgtcttga agaattttcc tgcattgctg agcacagggg
     3001 agttcttgaa actccctttt gagcgtcttg ccttcgtgct ttccagtaat agccttaagc
     3061 actgtactga acttgagctc tttaaggcta cctgtcgttg gcttcgcctg gaagagcctc
     3121 ggatggactt tgctgcaaaa ttaatgaaga acatacgatt tccactgatg acaccacagg
     3181 agctcattaa ttacgtgcaa acggtggatt tcatgagaac tgacaatact tgtgtgaatt
     3241 tgcttttgga agccagcaat taccaaatga tgccatatat gcagccagtt atgcagtcag
     3301 acaggactgc cattaggtct gacaccactc acttggttac actaggagga gtgctgaggc
     3361 agcagctggt tgtcagtaag gaattgcgca tgtatgatga aaaggcccat gagtggaaat
     3421 cgttagcccc catggatgcc ccaaggtacc agcatggcat cgccgtcatt ggaaattttc
     3481 tctatgtggt tggcggacag agtaattatg atacaaaagg aaaaacggca gttgatacag
     3541 tcttcagatt tgatcctcga tacaataaat ggatgcaagt tgcatcttta aatgaaaagc
     3601 gcaccttctt ccacctaagt gccctcaaag gatatctgta tgcagttggt gggcgaaatg
     3661 cagcaggtga actgcccaca gtagaatgtt acaatccaag aacaaatgaa tggacctatg
     3721 ttgccaaaat gagtgagccc cactatggcc atgctggaac tgtgtatgga ggagtgatgt
     3781 atatttcagg aggaattact catgatactt tccaaaagga gctcatgtgc tttgaccctg
     3841 atactgacaa atggatccag aaggcgccaa tgaccactgt cagaggtctg cattgcatgt
     3901 gtacagtggg agaaaggctc tatgtcattg gtggcaatca cttcagagga acaagtgatt
     3961 atgatgatgt cctaagctgt gaatactatt cacctatcct tgaccagtgg accccaattg
     4021 ctgccatgtt aagagggcag agtgatgttg gggtcgctgt cttcgaaaat aaaatctatg
     4081 tggttggggg gtattcttgg aataatcgtt gtatggtaga gatagtgcag aaatatgatc
     4141 cagataaaga tgaatggcat aaggtttttg atctgccaga atcccttggt ggcattcgtg
     4201 cttgcacact cacagttttt ccaccagaag aaaccacacc atcaccttct agagagtccc
     4261 ctctttctgc acctggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     4321 tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat cgctccggtg
     4381 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     4441 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     4501 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     4561 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     4621 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     4681 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     4741 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     4801 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     4861 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat gaccgagtac
     4921 aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc
     4981 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag
     5041 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5101 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5161 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5221 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5281 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5341 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5401 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5461 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa
     5521 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5581 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5641 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5701 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5761 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5821 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5881 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5941 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6001 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6061 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc
     6121 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6181 agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg
     6241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6421 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6481 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6601 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg
     6661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6721 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca
     6781 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6841 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7621 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8941 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     9001 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     9061 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     9121 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     9181 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     9241 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     9301 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     9361 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
