LOCUS       V024230                 9853 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024230
VERSION     V024230
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"
     promoter        3329..3347
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             3369..4676
                     /label="FBXL20(NM_032875)"
                     /note="FBXL20(NM_032875)"
                     /gene="FBXL20"
     misc_feature    4685..5258
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5292..5690
                     /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    5705..6293
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6380..6613
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6685..6806
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6846..6981
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(7002..7020)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7030..7046)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      7188..7643
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7669..7773
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7774..8634
                     /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      8805..9393
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9681..9702
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9717..9747
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9755..9771
                     /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     9779..9795
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9816..9834
                     /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 cctctagata atacgactca ctatagggcc cgggttggat
     3361 ccgccaccat gaggagggac gtgaacggag tgaccaagag caggtttgag atgttctcaa
     3421 atagtgatga agctgtaatc aataaaaaac ttcccaaaga actcctgtta cggatatttt
     3481 cttttctaga tgttgttacc ctgtgccgct gtgctcaggt ctccagggcc tggaatgttc
     3541 tggctctgga tggcagtaac tggcagcgaa ttgacctatt tgatttccag agggatattg
     3601 agggccgagt agtggagaat atttcaaaac gatgtggggg ctttttacga aagttaagtc
     3661 ttcgtggatg tcttggagtg ggagacaatg cattaagaac ctttgcacaa aactgcagga
     3721 acattgaagt actgaatcta aatgggtgta caaagacaac agacgctaca tgtactagcc
     3781 ttagcaagtt ctgttccaaa ctcaggcacc ttgacttggc ttcctgtaca tcaataacaa
     3841 acatgtctct aaaagctctg agtgagggat gtccactgtt ggagcagttg aacatttcct
     3901 ggtgtgacca agtaaccaag gatggcattc aagcactagt gaggggctgt gggggtctca
     3961 aggccttatt cttaaaaggc tgcacgcagc tagaagatga agctctcaag tacataggtg
     4021 cacactgccc tgaactggtg actttgaact tgcagacttg cttgcaaatc acagatgaag
     4081 gtctcattac tatatgcaga gggtgccata agttacaatc cctttgtgcc tctggctgct
     4141 ccaacatcac agatgccatc ctgaatgctc taggtcagaa ctgcccacgg cttagaatat
     4201 tggaagtggc aagatgttct caattaacag atgtgggctt taccactcta gccaggaatt
     4261 gccatgaact tgaaaagatg gacctggaag agtgtgttca gataacagat agcacattaa
     4321 tccaactttc tatacactgt cctcgacttc aagtattgag tctgtctcac tgtgagctga
     4381 tcacagatga tggaattcgt cacctgggga atggggcctg cgcccatgac cagctggagg
     4441 tgattgagct ggacaactgc ccactaatca cagatgcatc cctggagcac ttgaagagct
     4501 gtcatagcct tgagcggata gaactctatg actgccagca aatcacacgg gctggaatca
     4561 agagactcag gacccattta cccaatatta aagtccacgc ctacttcgca cctgtcactc
     4621 cacccccatc agtagggggc agcagacagc gcttctgcag atgctgcatc atcctatgag
     4681 ctagcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     4741 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     4801 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     4861 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     4921 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     4981 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     5041 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     5101 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     5161 ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     5221 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacact agagatccac
     5281 cggtcgccac catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa
     5341 cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct
     5401 ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg
     5461 cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg
     5521 tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc
     5581 ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg
     5641 cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcggccgcgt
     5701 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5761 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5821 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5881 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5941 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6001 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6061 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     6121 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6181 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6241 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     6301 tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6361 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     6421 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6481 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6541 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6601 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6661 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6781 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6901 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6961 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     7021 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     7081 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     7141 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     7201 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7261 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7321 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7381 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7441 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7501 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     7561 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7621 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     7681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7861 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7921 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7981 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     8041 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     8101 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     8161 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8221 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8281 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     8341 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8401 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8461 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8521 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8581 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8641 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8701 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8761 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8821 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8881 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8941 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     9001 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     9061 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     9121 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9181 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9241 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9301 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9361 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9421 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9481 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9541 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9601 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     9661 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     9721 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     9781 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     9841 aagctggagc tgc
//
