LOCUS       V029018                 8223 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029018
VERSION     V029018
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        65..368
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        369..572
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     protein_bind    595..619
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             619..1122
                     /label="DESI1(NM_015704)"
                     /note="DESI1(NM_015704)"
                     /gene="DESI1"
     protein_bind    complement(1121..1145)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             1147..1188
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     misc_feature    1230..1818
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             1890..2123
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    2201..2322
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      2362..2497
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(2518..2536)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(2546..2562)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      2704..3159
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3185..3289
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3290..4150
                     /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      4321..4909
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5197..5218
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5233..5263
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5271..5287
                     /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     5295..5311
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        5332..5350
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        5378..5604
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             5605..5785
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    5832..5957
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    6450..6683
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             6868..6912
                     /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"
     promoter        7087..7587
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             7599..7994
                     /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    8054..8171
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgtacaaaa aagttggcat ggagccgccg aatctctatc cggtgaagct ctacgtgtac
      661 gacctgtcca aaggcctggc ccggcggctc agccccatca tgctggggaa acaactggaa
      721 ggcatctggc acacatccat agttgtgcac aaggatgagt tcttcttcgg cagtggtggt
      781 atctccagct gccccccggg agggacattg cttgggcctc cagactctgt ggttgatgtg
      841 gggagtacag aagtcacaga agaaatcttt ctggagtacc tctcctccct gggggagtcc
      901 ctgttccgag gtgaggccta caacctcttt gaacacaatt gtaacacctt cagcaacgaa
      961 gtggcacagt tcctgactgg gcggaagatt ccttcttaca tcacagacct gccctctgaa
     1021 gttctctcca cgccctttgg acaggcactt cggcccctcc tggactccat tcagatccag
     1081 cctccaggag ggagctccgt gggcagaccc aacggccaga gcccaacttt cttgtacaaa
     1141 gtggttggta agcctatccc taaccctctc ctcggtctcg attctacgta gtaatgagct
     1201 agcgctaacc ggtggcgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa
     1261 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1321 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1381 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     1441 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     1501 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     1561 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     1621 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     1681 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     1741 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     1801 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
     1861 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     1921 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     1981 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     2041 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     2101 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
     2161 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     2221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2281 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     2341 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2401 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2461 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     2521 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2581 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2641 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2701 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3121 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     3181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4141 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4801 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4861 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4921 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4981 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5041 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5101 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5161 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     5221 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5281 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     5341 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     5401 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     5461 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     5521 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     5581 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     5641 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5701 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5761 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     5821 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     5881 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     5941 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     6001 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     6061 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     6121 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     6181 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     6241 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     6301 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     6361 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     6421 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     6481 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     6541 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     6601 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     6661 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     6721 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     6781 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     6841 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     6901 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     6961 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     7021 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     7081 ggacccttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
     7141 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
     7201 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
     7261 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
     7321 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
     7381 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
     7441 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
     7501 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
     7561 gaccgaatca ccgacctctc tccccagggg gtaccaccat ggccaagcct ttgtctcaag
     7621 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     7681 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     7741 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     7801 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     7861 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     7921 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     7981 atgtgtggga gggcctgcag ctgcagtagt aagaattcta gatcttgaga caaatggcag
     8041 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     8101 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     8161 ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc cggctcgagg
     8221 ggg
//
