LOCUS       V045127                 8888 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045127
VERSION     V045127
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..3813
                     /label="PNPLA2(NM_020376)"
                     /note="PNPLA2(NM_020376)"
                     /gene="PNPLA2"
     promoter        3858..4069
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4082..4350
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4377..4976
                     /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    4983..5571
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5645..5878
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5950..6071
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6090..6225
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6263..6279)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6287..6303
                     /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(6311..6341)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6356..6377
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6665..7253)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7424..8284)
                     /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(8285..8389)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8863..8879
                     /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 catgtttccc cgcgagaaga cgtggaacat ctcgttcgcg
     2341 ggctgcggct tcctcggcgt ctactacgtc ggcgtggcct cctgcctccg cgagcacgcg
     2401 cccttcctgg tggccaacgc cacgcacatc tacggcgcct cggccggggc gctcacggcc
     2461 acggcgctgg tcaccggggt ctgcctgggt gaggctggtg ccaagttcat tgaggtatct
     2521 aaagaggccc ggaagcggtt cctgggcccc ctgcacccct ccttcaacct ggtaaagatc
     2581 atccgcagtt tcctgctgaa ggtcctgcct gctgatagcc atgagcatgc cagtgggcgc
     2641 ctgggcatct ccctgacccg cgtgtcagac ggcgagaatg tcattatatc ccacttcaac
     2701 tccaaggacg agctcatcca ggccaatgtc tgcagcggtt tcatccccgt gtactgtggg
     2761 ctcatccctc cctccctcca gggggtgcgc tacgtggatg gtggcatttc agacaacctg
     2821 ccactctatg agcttaagaa caccatcaca gtgtccccct tctcgggcga gagtgacatc
     2881 tgtccgcagg acagctccac caacatccac gagctgcggg tcaccaacac cagcatccag
     2941 ttcaacctgc gcaacctcta ccgcctctcc aaggccctct tcccgccgga gcccctggtg
     3001 ctgcgagaga tgtgcaagca gggataccgg gatggcctgc gctttctgca gcggaacggc
     3061 ctcctgaacc ggcccaaccc cttgctggcg ttgccccccg cccgccccca cggcccagag
     3121 gacaaggacc aggcagtgga gagcgcccaa gcggaggatt actcgcagct gcccggagaa
     3181 gatcacatcc tggagcacct gcccgcccgg ctcaatgagg ccctgctgga ggcctgcgtg
     3241 gagcccacgg acctgctgac caccctctcc aacatgctgc ctgtgcgtct ggccacggcc
     3301 atgatggtgc cctacacgct gccgctggag agcgctctgt ccttcaccat ccgcttgctg
     3361 gagtggctgc ccgacgttcc cgaggacatc cggtggatga aggagcagac gggcagcatc
     3421 tgccagtacc tggtgatgcg cgccaagagg aagctgggca ggcacctgcc ctccaggctg
     3481 ccggagcagg tggagctgcg ccgcgtccag tcgctgccgt ccgtgccgct gtcctgcgcc
     3541 gcctacagag aggcactgcc cggctggatg cgcaacaacc tctcgctggg ggacgcgctg
     3601 gccaagtggg aggagtgcca gcgccagctg ctgctcggcc tcttctgcac caacgtggcc
     3661 ttcccgcccg aagctctgcg catgcgcgca cccgccgacc cggctcccgc ccccgcggac
     3721 ccagcatccc cgcagcacca gctggccggg cctgccccct tgctgagcac ccctgctccc
     3781 gaggcccggc ccgtgatcgg ggccctgggg ctgtgagcgg ccgcgaagga tctgcgatcg
     3841 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     3901 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     3961 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4021 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     4081 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     4141 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     4201 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     4261 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     4321 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga
     4381 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     4441 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     4501 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4561 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4621 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4681 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4741 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4801 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     4861 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4921 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg
     4981 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5041 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5101 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5161 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5221 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5281 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5341 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5401 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5461 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5521 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct
     5581 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     5641 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     5701 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     5761 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     5821 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     5881 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     5941 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6001 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6061 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     6121 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6181 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     6241 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6301 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6361 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6421 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6481 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7081 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7321 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7381 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7441 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7501 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7561 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7621 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7681 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7741 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7801 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7861 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7921 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7981 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8041 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8101 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8161 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8221 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8281 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8341 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8401 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     8461 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     8521 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     8581 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     8641 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     8701 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     8761 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     8821 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     8881 ccaagctg
//
