LOCUS       sequence-409856-        5910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid encoding photocaged SpyCatcher (pSC) fused to C-terminal 
            UnaG fragment. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5910)
  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 5910)
  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-cUnaG
FEATURES             Location/Qualifiers
     source          1..5910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     773..792
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(812..889)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1095..1114
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1198..1217
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1198..1216
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1217..1241
                     /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             1256..1278
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1289..1627
                     /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             1835..1852
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1905..1923)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1919..1966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2003..2458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2090..2109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2300..2321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2485..2589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2590..2658,2659..3450)
                     /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(2808..2827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3621..4209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4110..4129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4363..4380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4395..4537
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4463..4482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4623..4645
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4639..4830)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5639..5910,1..811))
                     /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"
ORIGIN
        1 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
       61 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
      121 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
      181 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
      241 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
      301 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
      361 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
      421 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
      481 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
      541 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
      601 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
      661 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
      721 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
      781 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
      841 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
      901 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
      961 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     1021 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     1081 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     1141 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     1201 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     1261 ttaactttaa gaaggagata tacatatggt tgatacctta tcaggtttat caagtgagca
     1321 aggtcagtcc ggtgatatga caattgaaga agatagtgct acccatatta aattctcata
     1381 gcgtgatgag gacggcaaag agttagctgg tgcaactatg gagttgcgtg attcatctgg
     1441 taaaactatt agtacatgga tttcagatgg acaagtgaaa gatttctacc tgtatccagg
     1501 aaaatataca tttgtcgaaa ccgcagcacc agacggttat gaggtagcaa ctgctattac
     1561 ctttacagtt aatgagcaag gtcaggttac tgtaaatggc aaagcaacta aaggtgacgc
     1621 tcatattggt ggcagtggtg gtgggagcgg cggtgttaaa tccgtagtca acttggtcgg
     1681 agaaaaactc gtatatgtcc aaaaatggga tggcaaagag acaacctacg tgcgtgaaat
     1741 taaagacggg aaactggttg tgacgctgac aatgggcgac gtggtggcag tgcgctcgta
     1801 ccggcgtgca accgaaaagc ttggtaccct cgagcaccac caccaccacc actgagatcc
     1861 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1921 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1981 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2041 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2101 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2161 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2221 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2281 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2341 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2401 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttca
     2461 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2521 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2581 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2641 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2701 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2761 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2821 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2881 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2941 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3001 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3061 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3121 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3181 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3241 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3301 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3361 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3421 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3481 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3541 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3601 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3661 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3721 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3781 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3841 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3901 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3961 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4021 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4081 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4141 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4201 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4261 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4321 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4381 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4441 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4501 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4561 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4621 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4681 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4741 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4801 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4861 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4921 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4981 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     5041 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     5101 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     5161 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     5221 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     5281 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     5341 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     5401 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     5461 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     5521 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5581 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     5641 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5701 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     5761 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     5821 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     5881 atgagctgtc ttcggtatcg tcgtatccca
//
