LOCUS       V029065                 9033 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029065
VERSION     V029065
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..1932
                     /label="USP27X(NM_001145073)"
                     /note="USP27X(NM_001145073)"
                     /gene="USP27X"
     protein_bind    complement(1931..1955)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             1957..1998
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     misc_feature    2040..2628
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             2700..2933
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    3011..3132
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      3172..3307
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(3328..3346)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3356..3372)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      3514..3969
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3995..4099
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4100..4960
                     /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      5131..5719
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6007..6028
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6043..6073
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6081..6097
                     /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     6105..6121
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        6142..6160
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        6188..6414
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             6415..6595
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    6642..6767
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    7260..7493
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             7678..7722
                     /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        7897..8397
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             8409..8804
                     /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    8864..8981
                     /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 gtgtaaggac tatgtatatg acaaagacat tgagcaaatt
      661 gccaaagaag agcaaggaga agctttgaaa ttacaagcct ccacctcaac agaggtttct
      721 caccagcagt gttcagtgcc aggccttggt gagaaattcc caacctggga aacaaccaaa
      781 ccagaattag aactgctggg gcacaacccg aggagaagaa gaatcacctc cagctttacg
      841 atcggtttaa gaggactcat caatcttggc aacacgtgct ttatgaactg cattgtccag
      901 gccctcaccc acacgccgat actgagagat ttctttctct ctgacaggca ccgatgtgag
      961 atgccgagtc ccgagttgtg tctggtctgt gagatgtcgt cgctgtttcg ggagttgtat
     1021 tctggaaacc cgtctcctca tgtgccctat aagttactgc acctggtgtg gatacatgcc
     1081 cgccatttag cagggtacag gcaacaggat gcccacgagt tcctcattgc agcgttagat
     1141 gtcctgcaca ggcactgcaa aggtgatgat gtcgggaagg cggccaacaa tcccaaccac
     1201 tgtaactgca tcatagacca aatcttcaca ggtggcctgc agtctgatgt cacctgtcaa
     1261 gcctgccatg gcgtctccac cacgatagac ccatgctggg acattagttt ggacttgcct
     1321 ggctcttgca cctccttctg gcccatgagc ccagggaggg agagcagtgt gaacggggaa
     1381 agccacatac caggaatcac caccctcacg gactgcttgc ggaggtttac gaggccagag
     1441 cacttaggaa gcagtgccaa aatcaaatgt ggtagttgcc aaagctacca ggaatctacc
     1501 aaacagctca caatgaataa attacctgtc gttgcctgtt ttcatttcaa acggtttgaa
     1561 cattcagcga aacagaggcg caagatcact acatacattt cctttcctct ggagctggat
     1621 atgacgccgt ttatggcctc aagtaaagag agcagaatga atggacaatt gcagctgcca
     1681 accaatagtg gaaacaacga aaataagtat tccttgtttg ctgtggttaa tcaccaagga
     1741 accttggaga gtggccacta taccagcttc atccggcacc acaaggacca gtggttcaag
     1801 tgtgatgatg ccgtcatcac taaggccagt attaaggacg tactggacag tgaagggtat
     1861 ttactgttct atcacaaaca ggtgctagaa catgagtcag aaaaagtgaa agaaatgaac
     1921 acacaagcct acccaacttt cttgtacaaa gtggttggta agcctatccc taaccctctc
     1981 ctcggtctcg attctacgta gtaatgagct agcgctaacc ggtggcgcgt taagtcgaca
     2041 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2101 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2161 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2221 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2281 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2341 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2401 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2461 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2521 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2581 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     2641 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     2701 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     2761 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     2821 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     2881 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta
     2941 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     3001 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3121 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     3181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3301 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     3361 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3421 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3481 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     3541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3721 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3841 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3961 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4081 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4561 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4681 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4741 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4801 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4861 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4921 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5881 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5941 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6001 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6061 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6121 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     6181 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     6241 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     6301 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     6361 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     6421 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6481 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6541 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     6601 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     6661 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     6721 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     6781 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     6841 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     6901 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     6961 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     7021 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7081 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     7141 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7201 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     7261 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     7321 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     7381 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     7441 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     7501 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     7561 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     7621 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     7681 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     7741 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     7801 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     7861 tcgtttcaga cccacctccc aaccccgagg ggacccttgc gccttttcca aggcagccct
     7921 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
     7981 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
     8041 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
     8101 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
     8161 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
     8221 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
     8281 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
     8341 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
     8401 gtaccaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
     8461 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
     8521 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
     8581 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
     8641 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
     8701 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
     8761 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggcctgcag ctgcagtagt
     8821 aagaattcta gatcttgaga caaatggcag tattcatcca caattttaaa agaaaagggg
     8881 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     8941 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     9001 gcagagatcc actttggcgc cggctcgagg ggg
//
