LOCUS       V035237                 8622 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035237
VERSION     V035237
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..3468
                     /label="ALKBH1(NM_006020)"
                     /note="ALKBH1(NM_006020)"
                     /gene="ALKBH1"
     CDS             3475..3540
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3584..3795
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3808..4076
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4111..4710
                     /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    4717..5305
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5379..5612
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5684..5805
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5824..5959
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5997..6013)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6021..6037
                     /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(6045..6075)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6090..6111
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6399..6987)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7158..8018)
                     /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(8019..8123)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8597..8613
                     /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 catggggaag atggcagcgg ccgtgggctc tgtggcgact
     2341 ctggcgactg agcccgggga ggacgccttt cggaaacttt tccgcttcta ccgtcagagc
     2401 cggcccggga ccgcagacct ggaaggggtc atcgacttct cggcggccca cgcagcccgt
     2461 ggcaagggtc ctggtgccca aaaggtgatc aaatctcagc taaatgtgtc ttctgtcagt
     2521 gagcagaatg catatagagc aggtcttcag cccgtcagca agtggcaagc ctatggactc
     2581 aaaggctatc ctgggtttat ttttatccca aaccccttcc tcccaggtta ccagtggcac
     2641 tgggtgaaac agtgccttaa gttatattcc cagaaaccta atgtatgtaa cctggacaaa
     2701 cacatgtcta aagaagagac ccaagatctg tgggaacaga gcaaagagtt cctgaggtat
     2761 aaagaagcga ctaaacggag accccgaagt ttactggaga aactgcgttg ggtgaccgta
     2821 ggctaccatt ataactggga cagtaagaaa tactcagcag atcattacac acctttccct
     2881 tctgacctgg gtttcctctc agagcaagta gccgctgcct gtggatttga ggatttccga
     2941 gctgaagcag ggatcctgaa ttactaccgc ctggactcca cactgggaat ccacgtagac
     3001 agatctgagc tagatcactc caaacccttg ctgtcattca gctttggaca gtccgccatc
     3061 tttctcctgg gtggtcttca aagggatgag gcccccacgg ccatgtttat gcacagtggt
     3121 gacatcatga taatgtcggg tttcagccgc ctcttgaacc acgcagtccc tcgtgtcctt
     3181 ccaaatccag aaggggaagg cctgcctcac tgcctagagg cacctctccc tgctgtcctc
     3241 ccgagagatt caatggtaga gccttgttct atggaggact ggcaggtgtg tgccagctac
     3301 ttgaagaccg ctcgtgttaa catgactgtc cgacaggtcc tggccacaga ccagaatttc
     3361 cctctagaac ccatcgagga tgaaaaaaga gacatcagta cagaaggttt ctgccatctg
     3421 gatgaccaga atagcgaagt aaaacgggcc aggataaacc ctgacagcgg atccgactac
     3481 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     3541 tgagcggccg caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     3601 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     3661 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     3721 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     3781 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     3841 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     3901 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     3961 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     4021 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     4081 ctgtgaccgg cgcctacccc gggcggccct atgaccgagt acaagcccac ggtgcgcctc
     4141 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4201 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4261 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4321 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4381 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4441 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4501 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     4561 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4621 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4681 tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt
     4741 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4801 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4861 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4921 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4981 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5041 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5101 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5161 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5221 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5281 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     5341 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     5401 acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     5461 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     5521 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     5581 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     5641 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     5701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     5821 gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5881 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5941 ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca
     6001 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6061 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6241 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6301 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7201 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7261 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7321 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7381 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7441 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7501 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7561 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7621 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7681 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7741 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7801 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7861 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7921 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7981 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8101 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8161 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     8221 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     8281 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     8341 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     8401 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     8461 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8521 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8581 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
