LOCUS       V042897                 9649 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042897
VERSION     V042897
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..4503
                     /label="CPXM1(NM_019609)"
                     /note="CPXM1(NM_019609)"
                     /gene="CPXM1"
     CDS             4510..4575
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4619..4830
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4843..5111
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5138..5737
                     /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    5744..6332
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6406..6639
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6711..6832
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6851..6986
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7024..7040)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7048..7064
                     /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(7072..7102)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7117..7138
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7426..8014)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8185..9045)
                     /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(9046..9150)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9624..9640
                     /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 catgtggggg ctcctgctcg ccctggccgc cttcgcgccg
     2341 gccgtcggcc cggctctggg ggcgcccagg aactcggtgc tgggcctcgc gcagcccggg
     2401 accaccaagg tcccaggctc gaccccggcc ctgcatagca gcccggcaca gccgccggcg
     2461 gagacagcta acgggacctc agaacagcat gtccggattc gagtcatcaa gaagaaaaag
     2521 gtcattatga agaagcggaa gaagctaact ctaactcgcc ccaccccact ggtgactgcc
     2581 gggccccttg tgacccccac tccagcaggg accctcgacc ccgctgagaa acaagaaaca
     2641 ggctgtcctc ctttgggtct ggagtccctg cgagtttcag atagccggct tgaggcatcc
     2701 agcagccagt cctttggtct tggaccacac cgaggacggc tcaacattca gtcaggcctg
     2761 gaggacggcg atctatatga tggagcctgg tgtgctgagg agcaggacgc cgatccatgg
     2821 tttcaggtgg acgctgggca ccccacccgc ttctcgggtg ttatcacaca gggcaggaac
     2881 tctgtctgga ggtatgactg ggtcacatca tacaaggtcc agttcagcaa tgacagtcgg
     2941 acctggtggg gaagtaggaa ccacagcagt gggatggacg cagtatttcc tgccaattca
     3001 gacccagaaa ctccagtgct gaacctcctg ccggagcccc aggtggcccg cttcattcgc
     3061 ctgctgcccc agacctggct ccagggaggc gcgccttgcc tccgggcaga gatcctggcc
     3121 tgcccagtct cagaccccaa tgacctattc cttgaggccc ctgcgtcggg atcctctgac
     3181 cctctagact ttcagcatca caattacaag gccatgagga agctgatgaa gcaggtacaa
     3241 gagcaatgcc ccaacatcac ccgcatctac agcattggga agagctacca gggcctgaag
     3301 ctgtatgtga tggaaatgtc ggacaagcct ggggagcatg agctggggga gcctgaggtg
     3361 cgctacgtgg ctggcatgca tgggaacgag gccctggggc gggagttgct tctgctcctg
     3421 atgcagttcc tgtgccatga gttcctgcga gggaacccac gggtgacccg gctgctctct
     3481 gagatgcgca ttcacctgct gccctccatg aaccctgatg gctatgagat cgcctaccac
     3541 cggggttcag agctggtggg ctgggccgag ggccgctgga acaaccagag catcgatctt
     3601 aaccataatt ttgctgacct caacacacca ctgtgggaag cacaggacga tgggaaggtg
     3661 ccccacatcg tccccaacca tcacctgcca ttgcccactt actacaccct gcccaatgcc
     3721 accgtggctc ctgaaacgcg ggcagtaatc aagtggatga agcggatccc ctttgtgcta
     3781 agtgccaacc tccacggggg tgagctcgtg gtgtcctacc cattcgacat gactcgcacc
     3841 ccgtgggctg cccgcgagct cacgcccaca ccagatgatg ctgtgtttcg ctggctcagc
     3901 actgtctatg ctggcagtaa tctggccatg caggacacca gccgccgacc ctgccacagc
     3961 caggacttct ccgtgcacgg caacatcatc aacggggctg actggcacac ggtccccggg
     4021 agcatgaatg acttcagcta cctacacacc aactgctttg aggtcactgt ggagctgtcc
     4081 tgtgacaagt tccctcacga gaatgaattg ccccaggagt gggagaacaa caaagacgcc
     4141 ctcctcacct acctggagca ggtgcgcatg ggcattgcag gagtggtgag ggacaaggac
     4201 acggagcttg ggattgctga cgctgtcatt gccgtggatg ggattaacca tgacgtgacc
     4261 acggcgtggg gcggggatta ttggcgtctg ctgaccccag gggactacat ggtgactgcc
     4321 agtgccgagg gctaccattc agtgacacgg aactgtcggg tcacctttga agagggcccc
     4381 ttcccctgca atttcgtgct caccaagact cccaaacaga ggctgcgcga gctgctggca
     4441 gctggggcca aggtgccccc ggaccttcgc aggcgcctgg agcggctaag gggacagaag
     4501 gatggatccg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     4561 gatgacgatg acaagtagcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg
     4621 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     4681 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4741 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4801 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     4861 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     4921 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     4981 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     5041 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     5101 cgttacagat ccaagctgtg accggcgcct acgctagatg accgagtaca agcccacggt
     5161 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     5221 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     5281 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     5341 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5401 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5461 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5521 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5581 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5641 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5701 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc gacaatcaac ctctggatta
     5761 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5821 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5881 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5941 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6001 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6061 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6121 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     6181 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6241 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6301 gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca
     6361 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     6421 actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     6481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     6661 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     6721 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6781 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6841 ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6901 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6961 gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc
     7021 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7081 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7141 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7201 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7261 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7321 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7861 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7981 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8041 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8101 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8161 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8221 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8281 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8341 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8401 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8461 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8521 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8581 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8641 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8701 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8761 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8821 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8881 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8941 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9001 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9061 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9121 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9181 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     9241 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     9301 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     9361 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     9421 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     9481 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     9541 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     9601 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
