LOCUS       V033313                10140 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033313
VERSION     V033313
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        9..235
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             236..416
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    463..588
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1081..1314
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1499..1543
                     /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    1810..1926
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2130..3312
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3335..4900
                     /label="FBXO7(NM_012179)"
                     /note="FBXO7(NM_012179)"
                     /gene="FBXO7"
     CDS             4901..4924
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     misc_feature    4972..5545
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5579..5977
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5992..6580
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6667..6900
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6972..7093
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7133..7268
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(7289..7307)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7317..7333)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      7475..7930
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7956..8060
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             8061..8921
                     /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"
     rep_origin      9092..9680
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9968..9989
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        10004..10034
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    10042..10058
                     /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"
     primer_bind     10066..10082
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        10103..10121
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
       61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
      121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
      181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
      541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
      601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
      661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
      721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
      781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
      841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
      901 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
      961 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1021 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1081 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1141 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1201 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1261 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1321 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1381 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     1441 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     1501 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     1561 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     1621 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     1681 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     1741 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     1801 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     1861 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa
     1921 aattttatcg atgcctcccc gtcaccaccc cccccaaccc gccccgaccg gagctgagag
     1981 taattcatac aaaaggactc gcccctgcct tggggaatcc cagggaccgt cgttaaactc
     2041 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc
     2101 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca
     2161 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     2221 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2281 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2341 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     2401 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt
     2461 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     2521 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     2581 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     2641 ttttgatgat atcctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     2701 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     2761 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     2821 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     2881 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     2941 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     3001 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     3061 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     3121 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     3181 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     3241 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     3301 caggtgtcgt gaaaactacc cctctagagc caccatgagg ctgcgggtgc ggcttctgaa
     3361 gcggacctgg ccgctggagg tgcccgagac ggagccgacg ctggggcatt tgcgctcgca
     3421 cctgaggcag tccctgctgt gcacctgggg gtacagttct aatacccgat ttacaattac
     3481 attgaactac aaggatcccc tcactggaga tgaagagacc ttggcttcat atgggattgt
     3541 ttctggggac ttgatatgtt tgattcttca agatgacatt ccagcgccta atataccttc
     3601 atccacagat tcagagcatt cttcactcca gaataatgag caaccctctt tggccaccag
     3661 ctccaatcag actagcatgc aggatgaaca accaagtgat tcattccaag gacaggcagc
     3721 ccagtctggt gtttggaatg acgacagtat gttagggcct agtcaaaatt ttgaagctga
     3781 gtcaattcaa gataatgcgc atatggcaga gggcacaggt ttctatccct cagaacccat
     3841 gctctgtagt gaatcggtgg aagggcaagt gccacattca ttagagacct tgtatcaatc
     3901 agctgactgt tctgatgcca atgatgcctt gatagtgttg atacatcttc tcatgttgga
     3961 gtcaggttac atacctcagg gcaccgaagc caaagcactg tccatgccgg agaagtggaa
     4021 gttgagcggg gtgtataagc tgcagtacat gcatcctctc tgcgagggca gctccgctac
     4081 tctcacctgt gtgcctttgg gaaacctgat tgttgtaaat gctacactaa aaatcaacaa
     4141 tgagattaga agtgtgaaaa gattgcagct gctaccagaa tcttttattt gcaaagagaa
     4201 actaggggaa aatgtagcca acatatacaa agatcttcag aaactctctc gcctctttaa
     4261 agaccagctg gtgtatcctc ttctggcttt tacccgacaa gcactgaacc taccagatgt
     4321 atttgggttg gtcgtcctcc cattggaact gaaactacgg atcttccgac ttctggatgt
     4381 tcgttccgtc ttgtctttgt ctgcggtttg tcgtgacctc tttactgctt caaatgaccc
     4441 actcctgtgg aggtttttat atctgcgtga ttttcgagac aatactgtca gagttcaaga
     4501 cacagattgg aaagaactgt acaggaagag gcacatacaa agaaaagaat ccccgaaagg
     4561 gcggtttgtg atgctcctgc catcgtcaac tcacaccatt ccattctatc ccaacccctt
     4621 gcaccctagg ccatttccta gctcccgcct tcctccagga attatcgggg gtgaatatga
     4681 ccaaagacca acacttccct atgttggaga cccaatcagt tcactcattc ctggtcctgg
     4741 ggagacgccc agccagtttc ctccactgag accacgcttt gatccagttg gcccacttcc
     4801 aggacctaac cccatcttgc cagggcgagg cggccccaat gacagatttc cctttagacc
     4861 cagcaggggt cggccaactg atggccggct gtcattcatg gattacaagg acgacgatga
     4921 caagtaggat ccacgcgtct ccggcctagg gataacaggg taatccgcta gcccctctcc
     4981 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     5041 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     5101 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     5161 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     5221 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     5281 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     5341 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     5401 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt
     5461 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     5521 tttcctttga aaaacacgat gataatatgg ccacactaga gatccaccgg tcgccaccat
     5581 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     5641 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     5701 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     5761 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     5821 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     5881 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     5941 tgaattgctg ccctctggtt atgtgtggga gggctaagcg gccgcgtcga caatcaacct
     6001 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     6061 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     6121 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     6181 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6241 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6301 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6361 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     6421 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6481 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     6541 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     6601 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6661 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     6721 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6781 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6841 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6901 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6961 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7021 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7081 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     7141 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     7201 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7261 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     7321 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     7381 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     7441 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     7501 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     7561 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     7621 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7681 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7741 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7801 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     7861 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     7921 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7981 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     8041 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     8101 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     8161 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     8221 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     8281 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     8341 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     8401 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     8461 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     8521 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     8581 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     8641 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     8701 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     8761 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     8821 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     8881 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     8941 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     9001 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     9061 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     9121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     9181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     9241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     9301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     9361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     9421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     9481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     9541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     9601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9721 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     9781 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     9841 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     9901 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     9961 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
    10021 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
    10081 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
//
