LOCUS       V038280                 8956 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038280
VERSION     V038280
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        join(8919..8956,1..473)
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             495..1094
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     LTR             1222..1455
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    1527..1648
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      1688..1823
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(1844..1862)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(1872..1888)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      2030..2485
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2511..2615
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             2616..3476
                     /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      3647..4235
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    4523..4544
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4559..4589
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4597..4613
                     /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     4621..4637
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        4658..4676
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        4704..4930
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             4931..5111
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    5158..5283
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    5776..6009
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             6194..6238
                     /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        6845..7048
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             7083..7895
                     /label="TMEM192(NM_001100389)"
                     /note="TMEM192(NM_001100389)"
                     /gene="TMEM192"
     CDS             7896..8570
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
                     /transl_table=1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
                     /label="mRFP1"
     CDS             8607..8627
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             8628..8708
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /transl_table=1
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
                     /label="3xHA"
     misc_feature    8753..8870
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
ORIGIN
        1 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
       61 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
      121 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
      181 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
      241 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
      301 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
      361 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
      421 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
      481 caccggagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
      541 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
      601 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
      661 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
      721 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
      781 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
      841 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
      901 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
      961 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     1021 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     1081 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     1141 cccatgcatc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     1201 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     1261 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     1321 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     1381 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     1441 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     1501 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     1561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1621 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     1681 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1741 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1801 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     1861 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1921 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1981 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     2041 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2101 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2161 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2221 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2281 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2341 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2401 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2461 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2701 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2881 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3001 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3121 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3181 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3481 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3781 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4261 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4321 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4381 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4441 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4501 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4561 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4621 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     4681 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
     4741 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
     4801 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
     4861 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
     4921 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4981 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5041 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     5101 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     5161 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     5221 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     5281 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     5341 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     5401 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     5461 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     5521 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     5581 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     5641 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     5701 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     5761 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     5821 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     5881 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     5941 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     6001 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     6061 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     6121 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     6181 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     6241 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     6301 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     6361 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     6421 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     6481 cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg
     6541 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     6601 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     6661 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgcc
     6721 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     6781 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     6841 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     6901 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     6961 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     7021 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
     7081 ccatggctgc tggaggcagg atggaggacg gttccttgga tatcacccag agtattgaag
     7141 acgacccact tctggatgcc cagcttctcc cacaccactc attacaagct cactttagac
     7201 cccgattcca tcctcttcct acagtcatca tagtgaatct tctgtggttt attcatctcg
     7261 tgtttgttgt tttagcattt ttaacaggtg tgctttgttc ttatcctaat ccaaatgagg
     7321 acaagtgccc aggaaattac acaaacccat tgaaagttca gacggttata atccttggga
     7381 aagttatttt gtggattctc catttactcc ttgaatgcta catccagtat caccacagca
     7441 aaatcagaaa ccgaggctat aacttgatct accgatcaac aaggcatctc aagagacttg
     7501 cgttgatgat acagtcctct ggcaacacag tgcttctcct catactgtgc atgcagcact
     7561 ccttcccaga gcctggcaga ttgtatcttg acctcattct ggccatcttg gcactggaac
     7621 tcatctgttc cctgatatgt ctcctcattt acacagtgaa aatccggaga tttaataaag
     7681 ctaaaccaga gcctgatata cttgaagaag aaaaaatcta tgcttacccc agcaatatta
     7741 cctcggagac tggattcaga actatttcaa gcctagaaga aattgttgaa aagcaaggag
     7801 acaccattga atacctgaag cgacacaatg cgctgctgag taagcgattg ttggctctca
     7861 cttcctcaga cctgggctgt cagccaagta gaacgatggc ctcctccgag gacgtcatca
     7921 aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga
     7981 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     8041 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc cagtacggct
     8101 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     8161 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     8221 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     8281 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     8341 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gatgaggctg aagctgaagg
     8401 acggcggcca ctacgacgcc gaggtcaaga ccacctacat ggccaagaag cccgtgcagc
     8461 tgcccggcgc ctacaagacc gacatcaagc tggacatcac ctcccacaac gaggactaca
     8521 ccatcgtgga acagtacgag cgcgccgagg gccgccactc caccggcgcc ggtggaggta
     8581 gtggaggcgg gagtgggggt ggaagcgaga acctgtactt ccaatcctac ccatacgatg
     8641 ttccagatta cgcttatccg tatgacgtcc ctgactatgc atacccctac gatgtacccg
     8701 attacgccta agaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa
     8761 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     8821 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     8881 acagggacag cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc
     8941 caaggcagcc ctgggt
//
