LOCUS       V031731                 9652 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031731
VERSION     V031731
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..4506
                     /label="DGKA(NM_001351033)"
                     /note="DGKA(NM_001351033)"
                     /gene="DGKA"
     CDS             4513..4578
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4622..4833
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4846..5114
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5141..5740
                     /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    5747..6335
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6409..6642
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6714..6835
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6854..6989
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7027..7043)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7051..7067
                     /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(7075..7105)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7120..7141
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7429..8017)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8188..9048)
                     /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(9049..9153)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9627..9643
                     /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 catggccaag gagaggggcc taataagccc cagtgatttt
     2341 gcccagctgc aaaaatacat ggaatactcc accaaaaagg tcagtgatgt cctaaagctc
     2401 ttcgaggatg gcgagatggc taaatatgtc caaggagatg ccattgggta cgagggattc
     2461 cagcaattcc tgaaaatcta tctcgaagtg gataatgttc ccagacacct aagcctggca
     2521 ctgtttcaat cctttgagac tggtcactgc ttaaatgaga caaatgtgac aaaagatgtg
     2581 gtgtgtctca atgatgtttc ctgctacttt tcccttctgg agggtggtcg gccagaagac
     2641 aagttagaat tcaccttcaa gctgtacgac acggacagaa atgggatcct ggacagctca
     2701 gaagtggaca aaattatcct acagatgatg cgagtggctg aatacctgga ttgggatgtg
     2761 tctgagctga ggccgattct tcaggagatg atgaaagaga ttgactatga tggcagtggc
     2821 tctgtctctc aagctgagtg ggtccgggct ggggccacca ccgtgccact gctagtgctg
     2881 ctgggtctgg agatgactct gaaggacgac ggacagcaca tgtggaggcc caagaggttc
     2941 cccagaccag tctactgcaa tctgtgcgag tcaagcattg gtcttggcaa acagggactg
     3001 agctgtaacc tctgtaagta cactgttcac gaccagtgtg ccatgaaagc cctgccttgt
     3061 gaagtcagca cctatgccaa gtctcggaag gacattggtg tccaatcaca tgtgtgggtg
     3121 cgaggaggct gtgagtccgg gcgctgcgac cgctgtcaga aaaagatccg gatctaccac
     3181 agtctgaccg ggctgcattg tgtatggtgc cacctagaga tccacgatga ctgcctgcaa
     3241 gcggtgggcc atgagtgtga ctgtgggctg ctccgggatc acatcctgcc tccatcttcc
     3301 atctatccca gtgtcctggc ctctggaccg gatcgtaaaa atagcaaaac aagccagaag
     3361 accatggatg atttaaattt gagcacctct gaggctctgc ggattgaccc tgttcctaac
     3421 acccacccac ttctcgtctt tgtcaatcct aagagtggcg ggaagcaggg gcaaagggtg
     3481 ctctggaagt tccagtatat attaaaccct cgacaggtgt tcaacctcct aaaggatggt
     3541 cctgagatag ggctccgatt attcaaggat gttcctgata gccggatttt ggtgtgtggt
     3601 ggagacggca cagtaggctg gattctagag accattgaca aagctaactt gccagttttg
     3661 cctcctgttg ctgtgttgcc cctgggtact ggaaatgatc tggctcgatg cctaagatgg
     3721 ggaggaggtt atgaaggaca gaatctggca aagatcctca aggatttaga gatgagtaaa
     3781 gtggtacata tggatcgatg gtctgtggag gtgatacctc aacaaactga agaaaaaagt
     3841 gacccagtcc cctttcaaat catcaataac tacttctcta ttggcgtgga tgcctctatt
     3901 gctcatcgat tccacatcat gcgagagaaa tatccggaga agttcaacag cagaatgaag
     3961 aacaagctat ggtacttcga atttgccaca tctgaatcca tcttctcaac atgcaaaaag
     4021 ctggaggagt ctttgacagt tgagatctgt gggaaaccgc tggatctgag caacctgtcc
     4081 ctagaaggca tcgcagtgct aaacatccct agcatgcatg gtggctccaa cctctggggt
     4141 gataccagga gaccccatgg ggatatctat gggatcaacc aggccttagg tgctacagct
     4201 aaagtcatca ccgaccctga tatcctgaaa acctgtgtac cagacctaag tgacaagaga
     4261 ctggaagtgg ttgggctgga gggtgcaatt gagatgggcc aaatctatac caagctcaag
     4321 aatgctggac gtcggctggc caagtgctct gagatcacct tccacaccac aaaaaccctt
     4381 cccatgcaaa ttgacggaga accctggatg cagacgccct gtacaatcaa gatcacccac
     4441 aagaaccaga tgcccatgct catgggccca cccccccgct ccaccaattt ctttggcttc
     4501 ttgagcggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     4561 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4621 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4681 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4741 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4801 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4861 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4921 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4981 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     5041 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     5101 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     5161 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     5221 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     5281 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5341 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5401 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5461 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5521 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5581 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5641 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5701 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5761 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5821 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5881 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5941 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6001 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6061 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6121 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     6181 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6241 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6301 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     6361 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6421 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     6481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6661 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6721 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6781 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6841 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6901 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6961 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     7021 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7081 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7141 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7201 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7261 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7621 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7861 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8101 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8161 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8221 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8281 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8341 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8401 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8461 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8521 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8581 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8641 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8701 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8761 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8821 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8881 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8941 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9001 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9061 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9181 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     9241 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     9301 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     9361 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     9421 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9481 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9541 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9601 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
