LOCUS       V022309                 9152 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022309
VERSION     V022309
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..4077
                     /label="Igf2bp2(NM_183029)"
                     /note="Igf2bp2(NM_183029)"
                     /gene="Igf2bp2"
     promoter        4122..4333
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4346..4614
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4641..5240
                     /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    5247..5835
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5909..6142
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6214..6335
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6354..6489
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6527..6543)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6551..6567
                     /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(6575..6605)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6620..6641
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6929..7517)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7688..8548)
                     /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(8549..8653)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9127..9143
                     /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 catgatgaac aagctgtaca ttgggaacct gagccccgcc
     2341 gtcaccgccg acgacctccg gcagctcttc ggggacagga agctgcccct ggcgggacag
     2401 gtcctactca agtccggcta cgccttcgtg gactaccccg accagaactg ggccatccgc
     2461 gccatcgaga ccctctcggg taaagtggaa ttgcatggga aaatcatgga agttgactac
     2521 tcagtctcta aaaagctaag gagcagaaga atccagattc ggaacatccc gcctcacctg
     2581 cagtgggagg tgttggatgg gctgttggct gaatatggga cagtggagaa cgtggagcaa
     2641 gtcaacacag atacagaaac tgccgttgtc aacgtcacct atatgacaag agaagaggca
     2701 aagctagcta ttgagaagct cagtgggcat cagtttgagg actactcctt caagatttcc
     2761 tacatccccg atgaagaggt gagctctcct tcaccccctc atcgtgcccg ggaacaaggc
     2821 cacggccccg ggagctcttc tcaggccaga cagattgatt tcccgctgcg gatcctggtc
     2881 cccacccagt ttgttggtgc catcatcgga aaggagggct tgaccataaa gaacatcact
     2941 aagcagaccc agtcccgggt agacatccac agaaaggaga actctggggc tgcagagaag
     3001 cctgtcacaa tccatgctac cccagaaggg acatctgaag catgccgcat gattcttgag
     3061 attatgcaaa aagaagctga tgagaccaaa ctggctgagg aggttcctct gaaaatcctg
     3121 gcccacaatg gcttcgttgg aagactgatt ggcaaagaag gcagaaacct gaagaaaata
     3181 gaacatgaga cagggaccaa gataaccatc tcatccttgc aggatttgag catttataac
     3241 cccgagagaa ccatcaccgt gaggggcacc attgaagcct gtgccaatgc tgagatagag
     3301 attatgaaga agctccgaga ggcctttgag aacgacatgc tggccgttaa ccaacaagcc
     3361 aatctgatcc cagggctaaa cctcagtgca cttggcatct tttcgactgg actgtctgtg
     3421 cttcctccac cagcagggcc ccgtggagtt ccccccagtc ctccctatca cccctttgct
     3481 acccactccg gatacttctc cagtctgtac cctcatcacc atttcggccc attcccacat
     3541 catcactcct acccagagca ggagactgta agtctcttca tcccaaccca ggctgtgggt
     3601 gctatcatcg ggaagaaggg ggcacacatc aaacagctcg ctcgatttgc tggtgcctcc
     3661 atcaagattg ctccagcaga aggtccagat gtcagtgaga ggatggtcat catcactggt
     3721 cctcctgaag cccagtttaa ggctcaggga cggatctttg ggaaactgaa ggaagaaaac
     3781 ttctttaatc ccaaagaaga agtgaagctg gaggcccaca tccgagtccc ctcgtcgacc
     3841 gctggccggg tgattggcaa gggcgggaaa accgtgaacg agctgcagaa cttgacaagt
     3901 gcagaagtta tcgtgcctcg tgaccaaacg ccagacgaga atgaggaagt gatcgtcaga
     3961 attatcgggc atttttttgc tagccagact gcacaacgca agatcaggga aattgtacag
     4021 caggtgaagc agcaggagca gagataccct cagggagtcg ccccacagcg cagcaagtga
     4081 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     4141 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     4201 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     4261 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     4321 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     4381 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     4441 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     4501 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     4561 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     4621 gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4681 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4741 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4801 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4861 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4921 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     4981 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5041 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5101 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5161 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5221 cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     5281 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5341 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5401 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5461 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5521 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5581 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5641 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5701 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5761 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5821 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     5881 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa
     5941 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6001 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6061 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6121 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6181 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6241 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6301 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6361 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6421 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6481 ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc
     6541 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6601 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6661 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6721 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6781 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6901 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7261 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7621 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7681 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7741 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7801 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7861 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7921 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7981 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8041 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8101 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8161 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8221 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8281 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8341 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8401 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8461 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8521 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8581 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8641 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     8701 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     8761 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     8821 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     8881 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     8941 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     9001 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     9061 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     9121 gacgttgtaa aacgacggcc agtgccaagc tg
//
