LOCUS       V030837                 9436 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030837
VERSION     V030837
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..4290
                     /label="ELF4(NM_001127197)"
                     /note="ELF4(NM_001127197)"
                     /gene="ELF4"
     CDS             4297..4362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4406..4617
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4630..4898
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4925..5524
                     /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    5531..6119
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6193..6426
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6498..6619
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6638..6773
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6811..6827)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6835..6851
                     /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(6859..6889)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6904..6925
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7213..7801)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7972..8832)
                     /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(8833..8937)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9411..9427
                     /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 catggctatt accctacagc ccagtgacct gatctttgag
     2341 ttcgcaagca acgggatgga tgatgatatc caccagctgg aagacccctc tgtgttccca
     2401 gctgtgatcg tggagcaggt accctaccct gatttactgc atctgtactc gggactggag
     2461 ttggacgacg ttcacaatgg catcataaca gacgggacct tgtgcatgac gcaggatcag
     2521 atcctggaag gcagtttttt gctgacagat gacaatgagg ccacctcgca caccatgtca
     2581 accgcggaag tcttactcaa tatggagtct cccagcgata tcctggatga gaagcagatc
     2641 ttcagtacct ccgaaatgct tccagactcg gaccctgcac cagctgtcac tctgcccaac
     2701 tacctgtttc ctgcctctga gcccgatgcc ctgaacaggg cgggtgacac tagtgaccag
     2761 gaggggcatt ctctggagga gaaggcctcc agagaggaaa gtgccaagaa gactgggaaa
     2821 tcaaagaaga gaatccggaa gaccaagggc aaccgaagta cctcacctgt cactgacccc
     2881 agcatcccca ttaggaagaa atcaaaggat ggcaaaggca gcaccatcta tctgtgggag
     2941 ttcctcctgg ctcttctgca agacagaaac acctgtccca agtacatcaa gtggacccag
     3001 cgagagaaag gcatcttcaa actggtggac tccaaagctg tgtccaagct gtgggggaag
     3061 cagaaaaaca agcctgacat gaactatgag acaatggggc gggcactaag atactactac
     3121 caaagaggca tactggccaa agtggaaggg cagaggctgg tgtaccagtt taaggagatg
     3181 cccaaggacc tggtggtcat tgaagatgag gatgagagca gcgaagccac agcagcccca
     3241 cctcaggcct ccacggcctc tgtggcctct gccagtacca cccggcgaac cagctccagg
     3301 gtctcatcca gatctgcccc ccagggcaag ggcagctctt cttgggagaa gccaaaaatt
     3361 cagcatgtcg gtctccagcc atctgcgagt ctggaattgg gaccgtcgct agacgaggag
     3421 atccccacta cctccaccat gctcgtctct ccagcagagg gccaggtcaa gctcaccaaa
     3481 gctgtgagtg catcttcagt gcccagcaac atccacctag gagtggcccc cgtggggtcg
     3541 ggctcggccc tgaccctgca gacgatccca ctgaccacgg tgctgaccaa tgggcctcct
     3601 gccagtacta ctgctcccac tcagctcgtt ctccagagtg ttccagcggc ctctactttc
     3661 aaggacacct tcactttgca ggcctctttc cccctgaacg ccagtttcca agacagccag
     3721 gtggcagccc caggggctcc actgattctc agtggcctcc cccaacttct ggctggggcc
     3781 aaccgtccga ccaacccggc gccacccacg gtcacagggg ctggaccagc agggcccagc
     3841 tctcagcccc ctgggactgt cattgctgcc ttcatcagga cttctggcac tacagcagcc
     3901 cctagggtca aggaggggcc actgaggtcc tcctcctatg ttcagggtat ggtgacgggg
     3961 gcccccatgg aggggctgct ggttcctgaa gagaccctga gggagctcct gagagatcag
     4021 gctcatcttc agccacttcc aacccaggtg gtttccaggg gttcccacaa tccgagcctt
     4081 ctgggcaacc agactttgtc tcctcccagc cgccccactg ttgggctgac cccagtggct
     4141 gaacttgagc tctcctcagg ctcagggtcc ctgctgatgg ctgagcctag tgtgaccaca
     4201 tctgggagcc ttctgacaag atcccccacc ccagcccctt tctccccatt caaccctact
     4261 tccctcatta agatggagcc ccatgacata ggatccgact acaaagacca tgacggtgat
     4321 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc
     4381 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4441 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     4501 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4561 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     4621 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     4681 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     4741 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     4801 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     4861 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     4921 ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     4981 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     5041 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5101 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5161 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5221 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5281 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5341 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5401 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5461 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5521 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5941 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6061 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6121 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6181 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     6241 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6301 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6361 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6421 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     6481 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6541 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6601 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     6661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6721 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     6781 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7021 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7081 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7621 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7981 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8041 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8101 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8161 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8221 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8281 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8341 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8401 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8461 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8521 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8581 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8641 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8701 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8761 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8821 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8881 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8941 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     9001 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     9061 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     9121 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     9181 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     9241 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     9301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9421 ggccagtgcc aagctg
//
