LOCUS       Exported                7090 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Vector for cotranslational expression of tagRFP-Bxb1.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-tagRFP-NLS-HA-Bxb1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7090)
  AUTHORS   Mulholland CB, Smets M, Schmidtmann E, Leidescher S, Markaki Y, 
            Hofweber M, Qin W, Manzo M, Kremmer E, Thanisch K, Bauer C, Rombaut 
            P, Herzog F, Leonhardt H, Bultmann S
  TITLE     A modular open platform for systematic functional studies under 
            physiological conditions.
  JOURNAL   Nucleic Acids Res. 2015 May 24. pii: gkv550.
  PUBMED    26007658
REFERENCE   2  (bases 1 to 7090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7090
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..664
                     /label=chicken beta-actin promoter
     intron          665..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1686..1704
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1710..2420
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             2436..2456
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2457..2483
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4009..4026)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4015..4239
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4311..4328)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4311..4327)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4320..4342)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4469..4924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4556..4575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4766..4787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4950..5054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5055..5915
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5273..5292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6086..6674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6575..6594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6828..6845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6962..6983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6998..7028
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7036..7052
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7041..7063
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7060..7076
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7060..7076
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
      541 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga
      841 aagccttaaa gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tgcggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctggcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcgggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccatctcc agcctcgggg ctgccgcagg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagatcctta
     1681 attaataata cgactcacta tagggggtca tggtgtctaa gggcgaagag ctgattaagg
     1741 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
     1801 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
     1861 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
     1921 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
     1981 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
     2041 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
     2101 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
     2161 tgtaccccgc tgacggcggc ctggaaggca gaagcgacat ggccctgaag ctcgtgggcg
     2221 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     2281 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     2341 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     2401 aactggggca caaacttaat cgacccgcca ccatgccaaa aaagaaaaga aaagtgtatc
     2461 cctatgatgt ccccgattat gccggttcaa gagccctggt cgtgattaga ctgagccgag
     2521 tgacagacgc caccacaagt cccgagagac agctggaatc atgccagcag ctctgtgctc
     2581 agcggggttg ggatgtggtc ggcgtggcag aggatctgga cgtgagcggg gccgtcgatc
     2641 cattcgacag aaagaggagg cccaacctgg caagatggct cgctttcgag gaacagccct
     2701 ttgatgtgat cgtcgcctac agagtggacc ggctgacccg ctcaattcga catctccagc
     2761 agctggtgca ttgggctgag gaccacaaga aactggtggt cagcgcaaca gaagcccact
     2821 tcgatactac cacacctttt gccgctgtgg tcatcgcact gatgggcact gtggcccaga
     2881 tggagctcga agctatcaag gagcgaaaca ggagcgcagc ccatttcaat attagggccg
     2941 gtaaatacag aggctccctg cccccttggg gatatctccc taccagggtg gatggggagt
     3001 ggagactggt gccagacccc gtccagagag agcggattct ggaagtgtac cacagagtgg
     3061 tcgataacca cgaaccactc catctggtgg cacacgacct gaatagacgc ggcgtgctct
     3121 ctccaaagga ttattttgct cagctgcagg gaagagagcc acagggaaga gaatggagtg
     3181 ctactgcact gaagagatct atgatcagtg aggctatgct gggttacgca acactcaatg
     3241 gcaaaactgt ccgggacgat gacggagccc ctctggtgag ggctgagcct attctcacca
     3301 gagagcagct cgaagctctg cgggcagaac tggtcaagac tagtcgcgcc aaacctgccg
     3361 tgagcacccc aagcctgctc ctgagggtgc tgttctgcgc cgtctgtgga gagccagcat
     3421 acaagtttgc cggcggaggg cgcaaacatc cccgctatcg atgcaggagc atggggttcc
     3481 ctaagcactg tggaaacggg acagtggcca tggctgagtg ggacgccttt tgcgaggaac
     3541 aggtgctgga tctcctgggt gacgctgagc ggctggaaaa agtgtgggtg gcaggatctg
     3601 actccgctgt ggagctggca gaagtcaatg ccgagctcgt ggatctgact tccctcatcg
     3661 gatctcctgc atatagagct gggtccccac agagagaagc tctggacgca cgaattgctg
     3721 cactcgctgc tagacaggag gaactggagg gcctggaggc caggccctct ggatgggagt
     3781 ggcgagaaac cggacagagg tttggggatt ggtggaggga gcaggacacc gcagccaaga
     3841 acacatggct gagatccatg aatgtccggc tcacattcga cgtgcgcggt ggcctgactc
     3901 gaaccatcga ttttggcgac ctgcaggagt atgaacagca cctgagactg gggtccgtgg
     3961 tcgaaagact gcacactggg atgtcctagg tcagagctcg ctgatcagcc tcgactgtgc
     4021 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4081 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4141 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4201 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4261 gctggggctc gagatccact agttctagcc tcgaggctag agcggccgcc actggccgtc
     4321 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4381 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4441 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4801 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4861 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4921 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4981 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5041 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5101 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5161 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5221 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5281 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5341 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5401 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5461 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5521 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5581 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5641 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5701 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5761 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5821 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5881 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5941 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6001 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6061 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6241 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6841 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6901 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6961 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7021 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7081 aggcgcgccg
//
