LOCUS       sequence-409853-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid encoding photocaged SpyCatcher (pSC) with a C-terminal 
            Sortase recognition sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  AUTHORS   Ruskowitz ER, Munoz-Robles BG, Strange AC, Butcher CH, Kurniawan S, 
            Filteau JR, DeForest CA
  TITLE     Spatiotemporal functional assembly of split protein pairs through a 
            light-activated SpyLigation.
  JOURNAL   Nat Chem. 2023 May;15(5):694-704. doi: 10.1038/s41557-023-01152-x. 
            Epub 2023 Apr 17.
  PUBMED    37069270
REFERENCE   2  (bases 1 to 5720)
  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: 
            "10.1038/s41557-023-01152-x"; journalName: "Nat Chem"; date: 
            "2023-05"; volume: "15"; issue: "5"; pages: "694-704"
COMMENT     Sequence Label: pSC-LPETG
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..342
                     /codon_start=1
                     /product="truncated version of the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyCatcher
                     /note="forms a covalent isopeptide bond with the SpyTag 
                     peptide (Zakeri et al., 2012)"
                     /translation="VDTLSGLSSEQGQSGDMTIEEDSATHIKFS*RDEDGKELAGATME
                     LRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG
                     KATKGDAHI"
     CDS             364..381
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(434..452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      448..495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      532..987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(619..638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     829..850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1013..1117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1118..1186,1187..1978)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1336..1355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2149..2737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2638..2657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2891..2908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2923..3062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2988..3007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3148..3170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3164..3355)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4164..5246)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5208..5227
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5247..5324)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5530..5549
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5633..5652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5633..5651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5652..5676
                     /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)."
     RBS             5691..5713
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atggttgata ccttatcagg tttatcaagt gagcaaggtc agtccggtga tatgacaatt
       61 gaagaagata gtgctaccca tattaaattc tcatagcgtg atgaggacgg caaagagtta
      121 gctggtgcaa ctatggagtt gcgtgattca tctggtaaaa ctattagtac atggatttca
      181 gatggacaag tgaaagattt ctacctgtat ccaggaaaat atacatttgt cgaaaccgca
      241 gcaccagacg gttatgaggt agcaactgct attaccttta cagttaatga gcaaggtcag
      301 gttactgtaa atggcaaagc aactaaaggt gacgctcata ttcttccaga aaccggcctc
      361 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
      421 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      481 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
      541 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      601 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      661 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      721 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
      781 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      841 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      901 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      961 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     1021 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1081 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1141 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1201 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1261 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1321 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1381 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1441 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1501 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1561 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1621 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1681 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1741 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1801 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1861 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1921 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1981 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2041 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2281 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2761 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2821 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2881 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2941 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     3001 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3061 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3121 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3181 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3241 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3301 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3361 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3421 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3481 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3541 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3601 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3661 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3721 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3781 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     3841 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3901 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3961 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4021 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4081 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4141 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4261 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4321 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4381 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4441 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4501 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4561 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4621 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4681 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4741 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4801 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4861 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4921 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4981 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     5041 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5101 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5161 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5221 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5281 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5341 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5401 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5461 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5521 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5581 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     5641 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     5701 tttaagaagg agatatacat
//
