LOCUS       V037681                 9460 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037681
VERSION     V037681
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..4314
                     /label="Prkg1(NM_001013833)"
                     /note="Prkg1(NM_001013833)"
                     /gene="Prkg1"
     CDS             4321..4386
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4430..4641
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4654..4922
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4949..5548
                     /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    5555..6143
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6217..6450
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6522..6643
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6662..6797
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6835..6851)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6859..6875
                     /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(6883..6913)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6928..6949
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7237..7825)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7996..8856)
                     /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(8857..8961)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9435..9451
                     /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 catgagcgaa ctggaggaag actttgcgaa gattctcatg
     2341 ctcaaggagg agaggatcaa agagctggag aagcggctgt cagagaagga ggaagaaatc
     2401 caggagctga agaggaaact ccacaaatgc cagtcggtgc tgccggtgcc ctccacccac
     2461 atcggcccgc ggaccacccg ggcgcagggc atctccgcgg agccgcagac ctacaggtcc
     2521 ttccacgacc tgcgccaggc gttccggaag ttcactaaat ccgagaggtc gaaggatctc
     2581 ataaaggagg ctatccttga caatgacttc atgaagaact tggagctgtc acagatccag
     2641 gagattgtgg actgtatgta ccccgtggaa tacggcaagg acagctgcat catcaaggaa
     2701 ggcgatgtgg ggtcactggt gtacgtcatg gaagatggga aggttgaagt cacaaaagaa
     2761 ggcgtgaagc tctgcaccat gggtcctgga aaagtgttcg gggagctggc tatactttac
     2821 aactgtaccc ggacagcgac cgtcaagact cttgtaaatg tgaaactctg ggccatcgat
     2881 cgacaatgtt ttcaaacaat aatgatgagg acaggactca tcaagcatac cgagtacatg
     2941 gaatttttaa aaagtgttcc aacattccag agccttcctg atgaaatcct cagcaagctg
     3001 gctgatgtcc tcgaagagac ccactatgaa aatggagaat atatcatcag gcagggtgca
     3061 agaggagaca ccttcttcat catcagtaaa gggcaggtga atgttactcg agaagactca
     3121 ccaagtgaag acccagtctt ccttagaact ttagggaagg gagattggtt tggagagaaa
     3181 gcgttgcagg gggaggatgt gagaacagca aatgttattg ccgcagaagc tgtcacctgc
     3241 cttgtgattg acagagactc tttcaagcat ttgattggag gactggacga tgtttctaac
     3301 aaagcatatg aggacgcaga agcaaaagca aaatatgaag ccgaagctgc cttcttcgcc
     3361 aacctgaagc tgtctgattt caacatcatt gacacccttg gagttggagg tttcggacga
     3421 gtagagctgg tccagttgaa aagtgaagaa tccaaaacat ttgcaatgaa gatcctcaag
     3481 aaacgccaca ttgtggacac cagacagcag gagcacatcc gctcagagaa gcagatcatg
     3541 cagggggctc attctgactt cattgtgagg ctgtacagga catttaaaga cagcaaatac
     3601 ttgtatatgt tgatggaagc gtgcctgggt ggagagctct ggactattct cagggatagg
     3661 ggttcgtttg aagactcaac aaccaggttt tacactgcat gtgtggtaga agcattcgcc
     3721 tatctgcatt ccaaaggaat catttacagg gacctcaagc cggagaatct catcctagat
     3781 catcgaggct atgccaaact ggttgacttt ggctttgcaa agaaaatagg atttggaaag
     3841 aaaacatgga ctttttgtgg gactccagaa tatgtagccc cagagatcat cctgaacaaa
     3901 ggccatgaca tttcagctga ctactggtca ctaggaattc tgatgtatga gcttctgact
     3961 ggcagcccac ctttctcagg cccagatcca atgaaaacct acaatatcat actgcggggg
     4021 attgacatga tagaatttcc aaagaagatt gcaaaaaatg ctgctaattt aattaaaaaa
     4081 ctatgcaggg acaacccatc agagaggtta ggaaatttga aaaacggagt caaagacatt
     4141 cagaaacaca agtggtttga gggctttaat tgggaaggct taagaaaagg caccttgaca
     4201 cctcccataa ttccaagtgt tgcgtcaccc acagacacaa gcaattttga cagtttccct
     4261 gaggacagcg atgagccacc acctgatgac aactcaggct gggacataga cttcggatcc
     4321 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4381 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4441 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4501 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4561 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4621 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4681 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4741 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4801 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4861 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4921 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     4981 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5041 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5101 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5161 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5221 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5281 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5341 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5401 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5461 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5521 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     5581 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5641 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5701 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5761 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5821 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5881 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5941 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6001 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6061 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6121 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6181 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6241 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6301 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6361 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6421 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6481 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6541 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6661 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6721 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6781 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9001 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     9061 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9121 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9181 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9241 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9301 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9361 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9421 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
