LOCUS       V044240                 9632 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044240
VERSION     V044240
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..3192
                     /codon_start=1
                     /product="modified&nbsp;bacterial dehalogenase that forms
                     covalent bonds with&nbsp;chloroalkane derivatives"
                     /transl_table=1
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
                     /label="HaloTag®"
     CDS             3226..4551
                     /label="CRBN(NM_016302)"
                     /note="CRBN(NM_016302)"
                     /gene="CRBN"
     promoter        4602..4813
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4826..5094
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5121..5720
                     /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    5727..6315
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6389..6622
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6694..6815
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6834..6969
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7007..7023)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7031..7047
                     /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(7055..7085)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7100..7121
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7409..7997)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8168..9028)
                     /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(9029..9133)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9607..9623
                     /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 catggcagaa atcggtactg gctttccatt cgacccccat
     2341 tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc
     2401 cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg
     2461 catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac
     2521 aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa
     2581 gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc
     2641 cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc
     2701 cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc
     2761 accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg
     2821 ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc
     2881 ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt
     2941 gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct
     3001 gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct
     3061 cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg
     3121 ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct gtctactctg
     3181 gagatttccg gaggaggggg aagcggaggc ggagggagtg aattcatggc cggcgaagga
     3241 gatcagcagg acgctgcgca caacatgggc aaccacctgc cgctcctgcc tgcagagagt
     3301 gaggaagaag atgaaatgga agttgaagac caggatagta aagaagccaa aaaaccaaac
     3361 atcataaatt ttgacaccag tctgccgaca tcacatacat acctaggtgc tgatatggaa
     3421 gaatttcatg gcaggacttt gcacgatgac gacagctgtc aggtgattcc agttcttcca
     3481 caagtgatga tgatcctgat tcccggacag acattacctc ttcagctttt tcaccctcaa
     3541 gaagtcagta tggtgcggaa tttaattcag aaagatagaa cctttgctgt tcttgcatac
     3601 agcaatgtac aggaaaggga agcacagttt ggaacaacag cagagatata tgcctatcga
     3661 gaagaacagg attttggaat tgagatagtg aaagtgaaag caattggaag acaaaggttc
     3721 aaagtccttg agctaagaac acagtcagat ggaatccagc aagctaaagt gcaaattctt
     3781 cccgaatgtg tgttgccttc aaccatgtct gcagttcaat tagaatccct caataagtgc
     3841 cagatatttc cttcaaaacc tgtctcaaga gaagaccaat gttcatataa atggtggcag
     3901 aaataccaga agagaaagtt tcattgtgca aatctaactt catggcctcg ctggctgtat
     3961 tccttatatg atgctgagac cttaatggac agaatcaaga aacagctacg tgaatgggat
     4021 gaaaatctaa aagatgattc tcttccttca aatccaatag atttttctta cagagtagct
     4081 gcttgtcttc ctattgatga tgtattgaga attcagctcc ttaaaattgg cagtgctatc
     4141 cagcgacttc gctgtgaatt agacattatg aataaatgta cttccctttg ctgtaaacaa
     4201 tgtcaagaaa cagaaataac aaccaaaaat gaaatattca gtttatcctt atgtgggccg
     4261 atggcagctt atgtgaatcc tcatggatat gtgcatgaga cacttactgt gtataaggct
     4321 tgcaacttga atctgatagg ccggccttct acagaacaca gctggtttcc tgggtatgcc
     4381 tggactgttg cccagtgtaa gatctgtgca agccatattg gatggaagtt tacggccacc
     4441 aaaaaagaca tgtcacctca aaaattttgg ggcttaacgc gatctgctct gttgcccacg
     4501 atcccagaca ctgaagatga aataagtcca gacaaagtaa tactttgctt gtaaggatcc
     4561 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     4621 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     4681 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     4741 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     4801 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     4861 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     4921 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     4981 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     5041 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     5101 gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     5161 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     5221 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     5281 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     5341 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     5401 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     5461 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5521 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5581 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5641 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5701 cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     5761 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5821 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5881 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5941 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     6001 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     6061 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     6121 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6181 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6241 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     6301 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     6361 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa
     6421 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6481 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6541 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6601 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6661 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6721 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6781 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6841 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6901 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6961 ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc
     7021 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7081 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7141 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7201 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     9061 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9121 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     9181 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     9241 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     9301 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     9361 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     9421 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     9481 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     9541 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     9601 gacgttgtaa aacgacggcc agtgccaagc tg
//
