LOCUS       sequence-321961-        8337 bp    DNA     circular SYN 14-APR-2025
DEFINITION  TU for the constitutive expression of Blue Fluorescent Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8337)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 8337)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pDGB3 alpha1 P35S:BFP:T35S (GB3416)
FEATURES             Location/Qualifiers
     source          1..8337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1034..1735
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    1986..2010
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3311..3940
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4369..5442
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5508..5702
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5938..5960)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6046..6186
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6101..6120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6201..6218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6372..6960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6452..6471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7048..7863)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7048..7863)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7771..7790
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8296..8320
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcccctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
     1021 tattctagtc gaaatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     1081 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     1141 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     1201 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     1261 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     1321 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     1381 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     1441 aacactcggc tgggaggcct tcaccgaaac gctgtacccc gctgacggcg gcctggaagg
     1501 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     1561 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     1621 ctacagactg gaaagaatca aggaggccaa caacgaaacc tacgtcgagc agcacgaggt
     1681 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attgagcttc
     1741 ggccatgcta gagtccgcaa aaatcaccag tctctctcta caaatctatc tctctctatt
     1801 tttctccaga ataatgtgtg agtagttccc agataaggga attagggttc ttatagggtt
     1861 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
     1921 ctatcaataa aatttctaat tcctaaaacc aaaatccagt gacctcgctg tcatgagacg
     1981 aattctgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
     2041 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa
     2101 ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca
     2161 gtcggcttct gacgttcagt gcagccgtca tctgaaaacg acatgtcgca caagtcctaa
     2221 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt
     2281 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc
     2341 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa
     2401 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc
     2461 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg
     2521 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag
     2581 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc
     2641 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc
     2701 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt
     2761 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag
     2821 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac
     2881 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc
     2941 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag
     3001 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga
     3061 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ctctcaaccg
     3121 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca
     3181 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa
     3241 acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca
     3301 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac
     3361 catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga
     3421 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac
     3481 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga
     3541 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc
     3601 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga
     3661 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt
     3721 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc
     3781 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac
     3841 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt
     3901 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa
     3961 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct
     4021 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg
     4081 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag
     4141 ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag
     4201 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc
     4261 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc
     4321 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc
     4381 aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg
     4441 aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg
     4501 tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg
     4561 ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc
     4621 tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg
     4681 aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag
     4741 gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg
     4801 gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc
     4861 cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga
     4921 aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg
     4981 cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg
     5041 attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag
     5101 ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt
     5161 caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc
     5221 cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc
     5281 agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac
     5341 ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc
     5401 taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta
     5461 gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggac tctttcctgt ggatagcacg
     5521 tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag
     5581 ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat
     5641 ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca
     5701 taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg
     5761 cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg
     5821 gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac
     5881 cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     5941 ctgacacatg cagctcccgg tgacggtcac agcttgtctg taagcggatg ccgggagcag
     6001 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     6061 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     6121 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6181 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6241 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6301 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6361 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6421 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6481 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6541 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6601 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6661 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6721 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6781 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6841 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6901 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6961 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7021 gggattttgg tcatgcattc taggtgatta gaaaaactca tcgagcatca aatgaaactg
     7081 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     7141 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     7201 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     7261 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     7321 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     7381 aaccaaaccg ttattcattc gtgattgcgc ctgagcgagt cgaaatacgc gatcgctgtt
     7441 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     7501 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttccctgg
     7561 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     7621 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     7681 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     7741 gtagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     7801 agcatccatg ttggaattta atcgcggcct tgagcaagac gtttcccgtt gaatatggct
     7861 cataacagaa cttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag
     7921 ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac cttaaatacc
     7981 agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta tcgacggagc
     8041 cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac aatcaacatg
     8101 ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg ttcttccgaa
     8161 tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc tgacgccgtc
     8221 ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag
     8281 ctgttggctg gctggtggca ggatatattg tggtgtaaac ataacgaatt cgtctca
//
