LOCUS       sequence-436037-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  this is microTag (Y7I mutation in SsrA)-Src(D388R/Y529F) with 
            mCerulean3 at C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  AUTHORS   Liu B, Stone OJ, Pablo M, Herron JC, Nogueira AT, Dagliyan O, Grimm 
            JB, Lavis LD, Elston TC, Hahn KM
  TITLE     Biosensors based on peptide exposure show single molecule 
            conformations in live cells.
  JOURNAL   Cell. 2021 Oct 28;184(22):5670-5685.e23. doi: 
            10.1016/j.cell.2021.09.026. Epub 2021 Oct 11.
  PUBMED    34637702
REFERENCE   2  (bases 1 to 6314)
  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: 
            "10.1016/j.cell.2021.09.026"; journalName: "Cell"; date: 
            "2021-10-28- 28"; volume: "184"; issue: "22"; pages: "5670-5685.e23"
COMMENT     Sequence Label: Src(KD/YF)-SsrA(Y7I)-mCer3
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             122..916
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             122..916
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(176..195)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     786..805
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1104..1123)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1148..1195
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1524..2112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2013..2032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2287..2590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2591..2794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2745..2765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2831..2840
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4484..4486,4487..5197)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIKA
                     NFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4526..4547)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4787..4806)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5134..5155
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5328..5449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5365..5384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5419..5438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5456..5911)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5593..5614)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5805..5824
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5938..6042
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6044..6314,1..87)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6044..6235
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6067..6087)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6252..6314,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
       61 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
      121 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      181 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      241 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      301 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      361 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      421 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      481 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
      541 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
      601 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
      661 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
      721 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
      781 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
      841 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
      901 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
      961 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1021 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1081 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     1141 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     1201 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     1261 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     1321 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     1381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1441 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1501 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1561 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1621 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1981 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2041 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2101 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2161 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2221 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2281 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2341 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2401 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2461 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2521 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2581 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2641 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2701 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2761 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
     2821 gcttaattaa gccaccatgg gcagcaacaa gagcaagccc aaggacgcca gccagcggcg
     2881 ccgcagcctg gagccctcgg aaaacgtgca cggggcaggg ggcgccttcc cggcctcaca
     2941 gacaccgagc aagcccgcct ccgccgacgg ccaccgcggg cccagcgccg ccttcgtgcc
     3001 gcccgcggcc gagcccaagc tcttcggagg cttcaactcc tcggacaccg tcacctcccc
     3061 gcagagggcg gggcctctgg caggtggggt gaccaccttt gtggccctct atgactatga
     3121 gtcacggaca gagactgacc tgtccttcaa gaaaggggag cggctgcaga ttgttaacaa
     3181 cacggaggga gactggtggc tggcacactc gctgagcacg ggacagaccg gttacatccc
     3241 cagcaactat gtggcgccct ccgactccat ccaggctgag gagtggtact ttggcaagat
     3301 cactagacgg gaatcagagc ggctgctgct caacgccgag aacccgagag ggaccttcct
     3361 cgtgagggag agtgagacca caaaaggtgc ctactgcctc tctgtatccg acttcgacaa
     3421 tgccaagggc ctaaatgtga aacactacaa gatccgcaag ctggacagcg gcggtttcta
     3481 catcacctcc cgcacccagt tcaacagcct gcagcagctc gtggcttact actccaaaca
     3541 tgctgatggc ctgtgtcacc gcctcactac cgtatgtccc acatccaagc ctcagaccca
     3601 gggattggcc aaggatgcgt gggagatccc ccgggagtcc ctgcggctgg aggtcaagct
     3661 gggccagggt tgcttcggag aggtgtggat ggggacctgg aacggcacca cgagggttgc
     3721 catcaaaact ctgaagccag gcaccatgtc cccagaggcc ttcctgcagg aggcccaagt
     3781 catgaagaaa cttaggcacg agaaactggt gcagctgtat gctgtggtgt cggaagaacc
     3841 catttacatt gtgacagagt acatgaacaa ggggagtctg ctggactttc tcaaggggga
     3901 aacgggcaaa tatttgcggc taccccagct ggtggacatg tctgctcaga tcgcttcagg
     3961 catggcctat gtggagcgga tgaactatgt gcaccggcgc cttcgagccg ccaatatcct
     4021 agtaggggag aacctggtgt gcaaagtggc cgactttggg ttggcccggc tcatagaaga
     4081 caacgaatac acagcccggc aaggtgccaa attccccatc aagtggaccg cccctgaagc
     4141 tgctctgtac ggcaggttca ccatcaagtc ggatgtgtgg tcctttggga ttctgctgac
     4201 cgagctcacc actaagggaa gagtgcccta tcctgggatg gtgaaccgtg aggttctgga
     4261 ccaggtggag cggggctacc ggatgccttg tccccccgag tgccccgagt ccctgcatga
     4321 ccttatgtgc cagtgctggc ggaaggagcc cgaggagcgg cccaccttcg agtacctgca
     4381 ggccttcctg gaagactact ttacgtccac tgagccacag ttccagcccg gggagaacgc
     4441 tgcaaatgat gaaaacatcc taggcggacc agggaagctt agcgtgagca agggcgagga
     4501 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     4561 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     4621 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgtcctg
     4681 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4741 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4801 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4861 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat
     4921 ccacggcaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcgg
     4981 cctcaactgc aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     5041 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     5101 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     5161 cgccgggatc actctcggca tggacgagct gtacaaggaa ttctaagcgg ccgcgactct
     5221 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5281 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5341 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5401 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     5461 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5521 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5581 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5641 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5701 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5761 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5821 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5881 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     5941 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6001 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     6061 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6121 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     6181 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     6241 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6301 gactaatttt tttt
//
