LOCUS       V020632                 8442 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020632
VERSION     V020632
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             2295..3296
                     /label="AMD1(NM_001634)"
                     /note="AMD1(NM_001634)"
                     /gene="AMD1"
     CDS             3303..3368
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3412..3623
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3636..3904
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3931..4530
                     /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    4537..5125
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5199..5432
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5504..5625
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5644..5779
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5817..5833)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5841..5857
                     /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(5865..5895)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5910..5931
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6978..7838)
                     /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(7839..7943)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8417..8433
                     /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 gagctagcgc caccatggaa gctgcacatt ttttcgaagg gaccgagaag ctgctggagg
     2341 tttggttctc ccggcagcag cccgacgcaa accaaggatc tggggatctt cgcactatcc
     2401 caagatctga gtgggacata cttttgaagg atgtgcaatg ttcaatcata agtgtgacaa
     2461 aaactgacaa gcaggaagct tatgtactca gtgagagtag catgtttgtc tccaagagac
     2521 gtttcatttt gaagacatgt ggtaccaccc tcttgctgaa agcactggtt cccctgttga
     2581 agcttgctag ggattacagt gggtttgact caattcaaag cttcttttat tctcgtaaga
     2641 atttcatgaa gccttctcac caagggtacc cacaccggaa tttccaggaa gaaatagagt
     2701 ttcttaatgc aattttccca aatggagcag catattgtat gggacgtatg aattctgact
     2761 gttggtactt atatactctg gatttcccag agagtcgggt aatcagtcag ccagatcaaa
     2821 ccttggaaat tctgatgagt gagcttgacc cagcagttat ggaccagttc tacatgaaag
     2881 atggtgttac tgcaaaggat gtcactcgtg agagtggaat tcgtgacctg ataccaggtt
     2941 ctgtcattga tgccacaatg ttcaatcctt gtgggtattc gatgaatgga atgaaatcgg
     3001 atggaactta ttggactatt cacatcactc cagaaccaga attttcttat gttagctttg
     3061 aaacaaactt aagtcagacc tcctatgatg acctgatcag gaaagttgta gaagtcttca
     3121 agccaggaaa atttgtgacc accttgtttg ttaatcagag ttctaaatgt cgcacagtgc
     3181 ttgcttcgcc ccagaagatt gaaggtttta agcgtcttga ttgccagagt gctatgttca
     3241 atgattacaa ttttgttttt accagttttg ctaagaagca gcaacaacag cagagtggat
     3301 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3361 atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc
     3421 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct
     3481 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     3541 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     3601 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg
     3661 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg
     3721 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac
     3781 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt
     3841 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca
     3901 gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc
     3961 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4021 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     4081 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4141 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4201 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4261 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4321 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     4381 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4441 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4501 tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt
     4561 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4621 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4681 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4741 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4801 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4861 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4921 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     4981 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5041 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5101 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     5161 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     5221 acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     5281 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     5341 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     5401 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     5461 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     5521 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5581 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     5641 gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5701 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5761 ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca
     5821 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5881 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7261 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7561 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7981 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     8041 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     8101 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     8161 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     8221 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     8281 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8341 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8401 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
