LOCUS       sequence-244066-        7932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon plasmid for stable expression of 
            mCerulean3-cpVenus[E172] FRET low control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7932)
  AUTHORS   Havranek O, Xu J, Kohrer S, Wang Z, Becker L, Comer JM, Henderson J,
            Ma W, Man Chun Ma J, Westin JR, Ghosh D, Shinners N, Sun L, Yi AF, 
            Karri AR, Burger JA, Zal T, Davis RE
  TITLE     Tonic B-cell receptor signaling in diffuse large B-cell lymphoma.
  JOURNAL   Blood. 2017 Aug 24;130(8):995-1006. doi: 
            10.1182/blood-2016-10-747303. Epub 2017 Jun 23.
  PUBMED    28646116
REFERENCE   2  (bases 1 to 7932)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1182/blood-2016-10-747303"; journalName: "Blood"; date: 
            "2017-08-24- 24"; volume: "130"; issue: "8"; pages: "995-1006"
COMMENT     Sequence Label: pSBbi-Pur-mCerulean3-spacer-cpVenus[E172]
FEATURES             Location/Qualifiers
     source          1..7932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1208..1217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1229..1246
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1250..1282
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1286..1294,1295..1309)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2744..3478
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     complement(3532..3549)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3538..3762
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4194..4298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4299..4367,4368..5159)
                     /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(4517..4536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5330..5918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5819..5838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6072..6089
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6463..6482
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6481..6536
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6582..6601
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6701..7300)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6784..6804)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7281..7300
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        join(7932,1..1182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccatactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt aaggtgtcgt gaaaactacc ccaagctggc
     1201 ctctgaggcc accatggtgc ggggttctca tcatcatcat catcatggta tggctagcat
     1261 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atcccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg agctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac gccatccacg gcaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1801 caaggccaac ttcggcctca actgcaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     2041 cagatcctgc gagagcctgg agaagaagac ggccactttt gagaacattg tctgcgtcct
     2101 gaaccgggag gtggagaggg tggccatgac tgccgaggcc tgcagccggc agcaccggct
     2161 ggaccaagac aagattgaag ccctgagtag caaggtgcag cagctggaga ggagcattgg
     2221 cctcaaggac ctggcgatgg ctgacttgga gcagaaggtc ttggagatgg aggcatccac
     2281 ctacgatggg gtcttcatct ggaagatctc agacttcgcc aggaagcgcc aggaagctgt
     2341 ggctggccgc atacccgcca tcttctcccc agccttctac accagcaggt acggctacaa
     2401 gatgtgtctg cgtatctacc tgaacggcga cggcaccggg cgaggaacac acctgtccct
     2461 cttctttgtg gtgatgaagg gcccgaatga cgccctgctg cggtggccct tcaaccagaa
     2521 ggtgacctta atgctgctcg accagaataa ccgggagcac gtgattgacg ccttcaggcc
     2581 cgacgtgact tcatcctctt ttcagaggcc agtcaacgac atgaacatcg caagcggctg
     2641 ccccctcttc tgccccgtct ccaagatgga ggcaaagaat tcctacgtgc gggacgatgc
     2701 catcttcatc aaggccattg tggacctgac agggctcgga tctatgggcg gcgtgcagct
     2761 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2821 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     2881 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2941 gggaggtacc ggtggatcta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3001 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3061 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     3121 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggccttcagt gcttcgcccg
     3181 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3241 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3301 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3361 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     3421 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagta
     3481 aggcctgtca ggccaagctt ggggatcaat tctctagagc tcgctgatca gcctcgactg
     3541 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3601 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3661 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3721 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3781 ccagctgggc ctactagata gcggacccct taccgaaaca tcgccgcatt ctgcagagga
     3841 gtcgaactta gtgtatgtaa acttctgacc cactggaatt gtgatacagt gaattataag
     3901 tgaaataatc tgtctgtaaa caattgttgg aaaaatgact tgtgtcatgc acaaagtaga
     3961 tgtcctaact gacttgccaa aactattgtt tgttaacaag aaatttgtgg agtagttgaa
     4021 aaacgagttt taatgactcc aacttaagtg tatgtaaact tccgacttca actgtatagg
     4081 gatccaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     4141 gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat gtgcgcggaa
     4201 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4261 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4321 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4381 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4441 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4501 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4561 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4621 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4681 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4741 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4801 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4861 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4921 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4981 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5041 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5101 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5161 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5221 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5281 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5341 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5401 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5461 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5521 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5581 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5641 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5701 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5761 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5821 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5881 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5941 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6001 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6061 cgaccgagcg cagcgagtca gtgagcgagg aagcggatcc ctatacagtt gaagtcggaa
     6121 gtttacatac accttagcca aatacattta aactcacttt ttcacaattc ctgacattta
     6181 atcctagtaa aaattccctg tcttaggtca gttaggatca ccactttatt ttaagaatgt
     6241 gaaatatcag aataatagta gagagaatga ttcatttcag cttttatttc tttcatcaca
     6301 ttcccagtgg gtcagaagtt tacatacact cgactcctct gcagaatgcg gcgatgtttc
     6361 ggtaaggggt cccgagtagc tagttcatgg cagccagcat atggcatatg ttgccaaact
     6421 ctaaaccaaa tactcattct gatgttttaa atgatttgcc ctcccatatg tccttccgag
     6481 tgagagacac aaaaaattcc aacacactat tgcaatgaaa ataaatttcc tttattagcc
     6541 agaagtcaga tgctcaaggg gcttcatgat gtccccataa tttttggcag agggaaaaag
     6601 atctcagtgg tatttgtgag ccagggcatt ggccacacca gccaccacct tctgataggc
     6661 agcctgcacc tgaggagtga attcacgaca ggccttcgaa tcaggcaccg ggcttgcggg
     6721 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
     6781 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
     6841 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
     6901 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     6961 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     7021 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     7081 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     7141 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc
     7201 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt
     7261 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtggcctca ggtgcagagg
     7321 tttctacagg gaaagagaga aaggctacag ttagtttgct gtgatgatgc taagtgccga
     7381 ggtcagatgt ctatggattg tacacggaat cccaacgcag gaaccaaaaa acactcaccc
     7441 aaaaaaatgg cgctaaggcc gagagaaagg cgaagttccg tctacggcta tataccttga
     7501 aacaggggcg gcagaagtga cgcaacgcgg ttgcgcggac ggcttcctgt gttttcattg
     7561 gtcgaagcgt tctgcgcgtg cgcagagatt ccccagggaa ggcgggccct cttccgaatg
     7621 tccaatccgg acattcttcc ggttggacct ccgcggtcag cgctgtgccc ggcgagagat
     7681 cacgtggggc gcggaggcgg tgctgctggg gcacggccgt ccagcctcgg cggccatatt
     7741 tttgaggggc tgttcatctc gttcacacgc tctgtccgcc atgtttgtga gtggaagcgc
     7801 cattacccct tcaagcgact gaaggctgca gggcctctgg tggcccgcat ggggagacca
     7861 gacccgccag gcccgccttt ccgcactcag tccgggctta ctttattttg tgagacaggg
     7921 tctcgcctag ag
//
