LOCUS       V029070                 8700 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029070
VERSION     V029070
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..1599
                     /label="COPS6(NM_006833)"
                     /note="COPS6(NM_006833)"
                     /gene="COPS6"
     protein_bind    complement(1598..1622)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             1624..1665
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     misc_feature    1707..2295
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             2367..2600
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    2678..2799
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      2839..2974
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(2995..3013)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3023..3039)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      3181..3636
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3662..3766
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3767..4627
                     /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      4798..5386
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5674..5695
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5710..5740
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5748..5764
                     /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     5772..5788
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        5809..5827
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        5855..6081
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             6082..6262
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    6309..6434
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    6927..7160
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             7345..7389
                     /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        7564..8064
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             8076..8471
                     /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    8531..8648
                     /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 ggcggcggcg gcggcggcgg ctgcagctac gaacgggacc
      661 ggaggaagca gcgggatgga ggtggatgca gcagtagtcc ccagcgtgat ggcctgcgga
      721 gtgactggga gtgtttccgt cgctctccat ccccttgtca ttctcaacat ctcagaccac
      781 tggatccgca tgcgctccca ggaggggcgg cctgtgcagg tgattggggc tctgattggc
      841 aagcaggagg gccgaaatat cgaggtgatg aactcctttg agctgctgtc ccacaccgtg
      901 gaagagaaga ttatcattga caaggaatat tattacacca aggaggagca gtttaaacag
      961 gtgttcaagg agctggagtt tctgggttgg tataccacag gggggccacc tgacccctcg
     1021 gacatccacg tccataagca ggtgtgtgag atcatcgaga gccccctctt tctgaagttg
     1081 aaccctatga ccaagcacac agatcttcct gtcagcgttt ttgagtctgt cattgatata
     1141 atcaatggag aggccacaat gctgtttgct gagctgacct acactctggc cacagaggaa
     1201 gcggaacgca ttggtgtaga ccacgtagcc cgaatgacag caacaggcag tggagagaac
     1261 tccactgtgg ctgaacacct gatagcacag cacagcgcca tcaagatgct gcacagccgc
     1321 gtcaagctca tcttggagta cgtcaaggcc tctgaagcgg gagaggtccc ctttaatcat
     1381 gagatcctgc gggaggccta tgctctgtgt cactgtctcc cggtgctcag cacagacaag
     1441 ttcaagacag atttttatga tcaatgcaac gacgtggggc tcatggccta cctcggcacc
     1501 atcaccaaaa cgtgcaacac catgaaccag tttgtgaaca agttcaatgt cctctacgac
     1561 cgacaaggca tcggcaggag aatgcgcggg ctctttttcc caactttctt gtacaaagtg
     1621 gttggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgtagta atgagctagc
     1681 gctaaccggt ggcgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     1741 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1801 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1861 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     1921 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     1981 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2041 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2101 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2161 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2221 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2281 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
     2341 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     2401 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     2461 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     2521 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     2581 agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt
     2641 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     2701 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2761 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     2821 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2881 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2941 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     3001 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3061 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3121 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     3181 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3241 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3301 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3361 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3421 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3481 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3541 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3601 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     3661 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3721 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3781 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3841 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3901 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3961 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4021 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4081 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4141 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4201 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4261 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4321 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4381 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4441 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4501 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4561 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4621 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4681 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4741 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4801 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4861 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4921 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4981 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5041 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5101 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5281 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5341 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5401 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5461 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5521 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     5581 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     5641 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     5701 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     5761 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     5821 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     5881 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     5941 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     6001 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     6061 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     6121 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6181 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6241 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     6301 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     6361 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     6421 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     6481 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     6541 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     6601 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     6661 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     6721 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     6781 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     6841 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     6901 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     6961 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     7021 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     7081 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     7141 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     7201 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     7261 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     7321 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     7381 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     7441 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     7501 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     7561 cccttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt
     7621 ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc
     7681 agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg
     7741 cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc
     7801 gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga
     7861 ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg
     7921 gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg
     7981 cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac
     8041 cgaatcaccg acctctctcc ccagggggta ccaccatggc caagcctttg tctcaagaag
     8101 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     8161 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     8221 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     8281 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     8341 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     8401 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     8461 tgtgggaggg cctgcagctg cagtagtaag aattctagat cttgagacaa atggcagtat
     8521 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     8581 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     8641 aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggggg
//
