LOCUS       V044370                 9105 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044370
VERSION     V044370
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..3823
                     /label="VKORC1(NM_024006)"
                     /note="VKORC1(NM_024006)"
                     /gene="VKORC1"
     CDS             3824..3889
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3937..4510
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4544..4942
                     /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    4957..5545
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5632..5865
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5937..6058
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6098..6233
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6254..6272)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6282..6298)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6440..6895
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6921..7025
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7026..7886
                     /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      8057..8645
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    8933..8954
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        8969..8999
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9007..9023
                     /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     9031..9047
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9068..9086
                     /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 caccatgggc agcacctggg ggagccctgg
     3361 ctgggtgcgg ctcgctcttt gcctgacggg cttagtgctc tcgctctacg cgctgcacgt
     3421 gaaggcggcg cgcgcccggg accgggatta ccgcgcgctc tgcgacgtgg gcaccgccat
     3481 cagctgttcg cgcgtcttct cctccaggtg gggcaggggt ttcgggctgg tggagcatgt
     3541 gctgggacag gacagcatcc tcaatcaatc caacagcata ttcggttgca tcttctacac
     3601 actacagcta ttgttaggtt gcctgcggac acgctgggcc tctgtcctga tgctgctgag
     3661 ctccctggtg tctctcgctg gttctgtcta cctggcctgg atcctgttct tcgtgctcta
     3721 tgatttctgc attgtttgta tcaccaccta tgctatcaac gtgagcctga tgtggctcag
     3781 tttccggaag gtccaagaac cccagggcaa ggctaagagg cacgactaca aagaccatga
     3841 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt aggatccacg
     3901 cgtctccggc ctagggataa cagggtaatc cgctagcccc tctccctccc ccccccctaa
     3961 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     4021 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     4081 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     4141 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     4201 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4261 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4321 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4381 accccattgt atgggatctg atctggggcc tcggtacaca tgctttacat gtgtttagtc
     4441 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4501 acgatgataa tatggccaca ctagagatcc accggtcgcc accatggcca agcctttgtc
     4561 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     4621 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     4681 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     4741 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     4801 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     4861 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     4921 tggttatgtg tgggagggct aagcggccgc gtcgacaatc aacctctgga ttacaaaatt
     4981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5101 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5161 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5221 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5281 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5341 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     5401 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5461 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5521 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga
     5581 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5641 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     5701 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5761 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5821 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     5881 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     5941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6061 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6121 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6181 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac
     6241 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     6301 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     6361 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     6421 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6481 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6541 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6601 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6661 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6721 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6781 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6841 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     6901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7081 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7141 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7201 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7261 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7321 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7381 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7441 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7501 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7561 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7621 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7681 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7741 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7801 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7861 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8701 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     8761 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8821 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8881 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     8941 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     9001 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     9061 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgc
//
