LOCUS       V041701                 9538 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041701
VERSION     V041701
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..4392
                     /label="ZNF418(NM_001317027)"
                     /note="ZNF418(NM_001317027)"
                     /gene="ZNF418"
     CDS             4399..4464
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4508..4719
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4732..5000
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5027..5626
                     /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    5633..6221
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6295..6528
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6600..6721
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6740..6875
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6913..6929)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6937..6953
                     /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(6961..6991)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7006..7027
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7315..7903)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8074..8934)
                     /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(8935..9039)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9513..9529
                     /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 catgcaagaa ggccttcata gaatgacttg ggactgcatt
     2341 gctattggga gccatgagaa ttctgtgcag ggcactgtgg catttgaaga tgtggctgtg
     2401 aacttttccc aggaggagtg gagtctcctt agtgaggttc agagatgcct ttaccatgac
     2461 gtgatgctgg agaactgggt acttatatcc tccctgggtt gttggtgtgg atcagaagat
     2521 gaggaggcac cttctaagaa gagcatttct atacaaagag tgtctcaggt cagcactcct
     2581 ggggcaggtg tgtctcccaa gaaggcccac tcttgtgaaa tgtgtggcgc gatcttggga
     2641 gacattttgc acttggcaga tcatcagggg acacatcaca agcagaaact gcacaggtgt
     2701 gaggcatggg ggaataaatt gtatgatagt tcaaaccgtc cgcaccagaa tcagtacctt
     2761 ggagagaaac cctatagaag cagtgttgag gaagcattgt ttgtgaagag gtgtaagttc
     2821 catgtgtcag aggagtcatc tatcttcatt cagagtggaa aggacttttt gcccagctca
     2881 ggattactgc tgcaggaggc cactcacact ggggagaagt caaacagcaa acctgagtgt
     2941 gagtctccct ttcagtgggg agatactcat tacagctgtg gagaatgcat gaaacattct
     3001 agcaccaaac acgtatttgt tcaacagcag agacttccct ctagagagga atgttattgc
     3061 tgggaatgtg ggaaatcctt tagcaaatat gatagcgtca gtaatcatca gagagttcac
     3121 actgggaaaa gaccttatga atgtggagaa tgtgggaaat cttttagtca taagggcagc
     3181 cttgttcagc atcagcgagt tcacactggg aaaagacctt atgaatgtgg agaatgtggg
     3241 aaatctttta gtcataaggg cagccttgtt cagcatcagc gagttcatac tggagaaaga
     3301 ccttatgagt gtggagaatg tgggaaatct tttagtcaaa atggtactct cattaaacat
     3361 caacgagttc acactggaga aagaccttat gagtgtgaag aatgtgggaa atgttttact
     3421 cagaagggca atctcattca acatcaacga ggtcacacta gtgaaagacc ttatgagtgt
     3481 gaagaatgtg gaaaatgttt tagtcaaaag ggcaccctaa ctgaacatca tcgagttcac
     3541 actagagaac gaccttatga gtgtggagaa tgtgggaaat cttttagtcg aaagggacac
     3601 cttaggaacc atcagcgagg tcacactgga gaaagacctt acgagtgtgg agaatgtggg
     3661 aaatctttta gtcgaaaggg caacctcatt cagcatcagc gaagccacac tggagaaagg
     3721 ccttatgagt gtagagagtg taggaaatta tttaggggca agtcccacct cattgaacac
     3781 cagagagttc acactggaga aaggccatat gaatgtaatg aatgtgggaa atcatttcaa
     3841 gacagctctg ggtttcgtgt tcatcagaga gttcacactg gagaaaaacc gtttgagtgt
     3901 agtgaatgtg ggaagtcatt tcctcaaagc tgttccctcc ttcgacatcg gagagttcat
     3961 actggagaaa ggccttatga atgtggagaa tgtggaaagt catttcatca gagctcttcc
     4021 ctccttcgac atcagaaaac tcacactgca gaaagacctt atgagtgcag agaatgtggg
     4081 aaattcttct ccagtctcct tgaacacagg agagttcaca ctggagaaag gccttatgaa
     4141 tgcagggaat gtggaaaaac atttactcga aggtctgcgc attttaaaca tcagagactt
     4201 catactcgag gaaagcctta cgagtgcagc gaatgtggga aatcctttgc tgaaaccttc
     4261 agtcttactg aacacaggag agtacacact ggagaaaggc cttatgagtg cagtgaatgt
     4321 ggaaaatcat ttcatcgaag ctcttctctc cttcgacatc agagagttca cacagaaaga
     4381 agtccttaca agggatccga ctacaaagac catgacggtg attataaaga tcatgacatc
     4441 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc
     4501 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     4561 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     4621 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4681 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     4741 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     4801 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     4861 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     4921 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     4981 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa
     5041 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     5101 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     5161 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5221 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5281 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5341 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5401 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5461 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5521 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5581 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc
     5641 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5701 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5761 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5821 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5881 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5941 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6001 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     6061 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6121 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6181 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     6241 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6301 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     6361 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6421 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6481 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6541 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     6601 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6661 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6721 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     6781 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6841 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     6901 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6961 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7021 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7741 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9061 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9121 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9181 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9241 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9301 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9361 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9421 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9481 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
