LOCUS       V035821                 9395 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035821
VERSION     V035821
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..4314
                     /label="RIPK1(NM_003804)"
                     /note="RIPK1(NM_003804)"
                     /gene="RIPK1"
     promoter        4365..4576
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4589..4857
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4884..5483
                     /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    5490..6078
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6152..6385
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6457..6578
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6597..6732
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6770..6786)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6794..6810
                     /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(6818..6848)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6863..6884
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7172..7760)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7931..8791)
                     /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(8792..8896)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9370..9386
                     /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 catgcaacca gacatgtcct tgaatgtcat taagatgaaa
     2341 tccagtgact tcctggagag tgcagaactg gacagcggag gctttgggaa ggtgtctctg
     2401 tgtttccaca gaacccaggg actcatgatc atgaaaacag tgtacaaggg gcccaactgc
     2461 attgagcaca acgaggccct cttggaggag gcgaagatga tgaacagact gagacacagc
     2521 cgggtggtga agctcctggg cgtcatcata gaggaaggga agtactccct ggtgatggag
     2581 tacatggaga agggcaacct gatgcacgtg ctgaaagccg agatgagtac tccgctttct
     2641 gtaaaaggaa ggataatttt ggaaatcatt gaaggaatgt gctacttaca tggaaaaggc
     2701 gtgatacaca aggacctgaa gcctgaaaat atccttgttg ataatgactt ccacattaag
     2761 atcgcagacc tcggccttgc ctcctttaag atgtggagca aactgaataa tgaagagcac
     2821 aatgagctga gggaagtgga cggcaccgct aagaagaatg gcggcaccct ctactacatg
     2881 gcgcccgagc acctgaatga cgtcaacgca aagcccacag agaagtcgga tgtgtacagc
     2941 tttgctgtag tactctgggc gatatttgca aataaggagc catatgaaaa tgctatctgt
     3001 gagcagcagt tgataatgtg cataaaatct gggaacaggc cagatgtgga tgacatcact
     3061 gagtactgcc caagagaaat tatcagtctc atgaagctct gctgggaagc gaatccggaa
     3121 gctcggccga catttcctgg cattgaagaa aaatttaggc ctttttattt aagtcaatta
     3181 gaagaaagtg tagaagagga cgtgaagagt ttaaagaaag agtattcaaa cgaaaatgca
     3241 gttgtgaaga gaatgcagtc tcttcaactt gattgtgtgg cagtaccttc aagccggtca
     3301 aattcagcca cagaacagcc tggttcactg cacagttccc agggacttgg gatgggtcct
     3361 gtggaggagt cctggtttgc tccttccctg gagcacccac aagaagagaa tgagcccagc
     3421 ctgcagagta aactccaaga cgaagccaac taccatcttt atggcagccg catggacagg
     3481 cagacgaaac agcagcccag acagaatgtg gcttacaaca gagaggagga aaggagacgc
     3541 agggtctccc atgacccttt tgcacagcaa agaccttacg agaattttca gaatacagag
     3601 ggaaaaggca ctgcttattc cagtgcagcc agtcatggta atgcagtgca ccagccctca
     3661 gggctcacca gccaacctca agtactgtat cagaacaatg gattatatag ctcacatggc
     3721 tttggaacaa gaccactgga tccaggaaca gcaggtccca gagtttggta caggccaatt
     3781 ccaagtcata tgcctagtct gcataatatc ccagtgcctg agaccaacta tctaggaaat
     3841 acacccacca tgccattcag ctccttgcca ccaacagatg aatctataaa atataccata
     3901 tacaatagta ctggcattca gattggagcc tacaattata tggagattgg tgggacgagt
     3961 tcatcactac tagacagcac aaatacgaac ttcaaagaag agccagctgc taagtaccaa
     4021 gctatctttg ataataccac tagtctgacg gataaacacc tggacccaat cagggaaaat
     4081 ctgggaaagc actggaaaaa ctgtgcccgt aaactgggct tcacacagtc tcagattgat
     4141 gaaattgacc atgactatga gcgagatgga ctgaaagaaa aggtttacca gatgctccaa
     4201 aagtgggtga tgagggaagg cataaaggga gccacggtgg ggaagctggc ccaggcgctc
     4261 caccagtgtt ccaggatcga ccttctgagc agcttgattt acgtcagcca gaactaagga
     4321 tccgcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     4381 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     4441 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     4501 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     4561 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     4621 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     4681 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     4741 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     4801 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa
     4861 gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     4921 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     4981 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5041 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5101 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5161 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5221 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5281 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5341 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5401 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5461 tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa
     5521 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5581 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5641 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5701 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5761 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5821 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5881 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5941 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6001 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6061 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     6121 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     6181 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6241 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6301 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6361 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6421 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6481 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6541 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6601 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6661 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6721 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     6781 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6841 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6901 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7981 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8041 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8101 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8161 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8221 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8281 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8341 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8401 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8461 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8521 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8581 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8641 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8701 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8761 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8821 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8881 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8941 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     9001 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     9061 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     9121 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     9181 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     9241 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     9301 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     9361 cacgacgttg taaaacgacg gccagtgcca agctg
//
