LOCUS       sequence-460063-        6248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to express cyclic sortase cW11 (recognizes APXTG motif) from E.
            coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6248)
  AUTHORS   Whedon SD, Lee K, Wang ZA, Zahn E, Lu C, Yapa Abeywardana M, Fairall
            L, Nam E, DuBois-Coyne S, De Ioannes P, Sheng X, Andrei A, Lundberg 
            E, Jiang J, Armache KJ, Zhao Y, Schwabe JWR, Wu M, Garcia BA, Cole 
            PA
  TITLE     Circular Engineered Sortase for Interrogating Histone H3 in 
            Chromatin.
  JOURNAL   J Am Chem Soc. 2024 Nov 25. doi: 10.1021/jacs.4c12585.
  PUBMED    39585806
REFERENCE   2  (bases 1 to 6248)
  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"; journalName: "J Am Chem 
            Soc. 2024 Nov 25. doi: 10.1021/jacs.4c12585."
COMMENT     Sequence Label: pSW031
FEATURES             Location/Qualifiers
     source          1..6248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     226..245
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(265..342)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      495..581
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     716..735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        716..734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    735..759
                     /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             774..796
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1383..1400
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1753..1908
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(1973..1991)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1987..2034
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2094..2198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2199..2267,2268..3059)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2417..2436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3101..3614)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3296..3317)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3508..3527
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3725..4313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4214..4233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4467..4484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4506..4525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4666..4688
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4682..4873)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5430..6248,1..264))
                     /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 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
       61 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
      121 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
      181 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
      241 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
      301 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
      361 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
      421 ccgccgcaag gaatggtgca tgccggcatg ccgccctttc gtcttcaaga attaattccc
      481 aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      541 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      601 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga
      661 attggggatc ggaattaatt cccggtttaa accggggatc tcgatcccgc gaaattaata
      721 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt
      781 aactttaaga aggagatata catatgatca aaatagccac acgtaaatat ttaggcaaac
      841 aaaatgtcta tgacattgga gttgagcgcg accataattt tgcactcaaa aatggcttca
      901 tagcttctaa ctgctttaac ggcggcagca gcatgcaagc taaacctcaa attccgaaag
      961 ataaatcgaa agtggcaggc tatattgaaa ttccagatgc tgatattaaa gaaccagtat
     1021 atccaggacc agcaacatct gaacaattaa atagaggtgt gagctttgca gaagaaaacc
     1081 agtctctaga cgaccagaat attgtgattg caggacacac tttcattgac cgtccgaact
     1141 accagttcac caacctgaag gcggcgaaga tgggctccat ggtgtacttc aaggtgggca
     1201 acgagacccg caagtacaag atgacctcca tccgcaacgt gaagccgcag gacgtgggca
     1261 tgcatcttgc ggagaagggc aaggacaagc agctgaccct gatcacctgc gacgactaca
     1321 acgaggaaac cggcgtgtgg gagacccgca agatcttcgt ggcgaccgag gtgaagctcg
     1381 agcaccacca ccaccaccac gcggaatatt gtttaagcta tgaaacggaa atattgacag
     1441 tagaatatgg attattaccg attggtaaaa ttgtagaaaa gcgcatcgaa tgtactgttt
     1501 atagcgttga taataatgga aatatttata cacaacctgt agcacaatgg cacgatcgcg
     1561 gagaacaaga ggtgtttgag tattgtttgg aagatggttc attgattcgg gcaacaaaag
     1621 accataagtt tatgactgtt gatggtcaaa tgttgccaat tgatgaaata tttgaacgtg
     1681 aattggattt gatgcgggtt gataatttgc cgaattaatt aaactggcct caccggtctg
     1741 aactcaggcc tcacgacaaa tcctggtgta tccgcttggc aggtcaacac agcttatact
     1801 gcgggacaat tggtcacata taacggcaag acgtataaat gtttgcagcc ccacacctcc
     1861 ttggcaggat gggaaccatc caacgttcct gccttgtggc agcttcaatg actgcaggaa
     1921 ggggatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1981 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     2041 ggaggaacta tatccggata actacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2101 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2161 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2221 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2281 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2341 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttc ccaatgatga
     2401 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     2461 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2521 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2581 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2641 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2701 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2761 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2821 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2881 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2941 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3001 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3061 tgtcagacca agtttactca tatatacttt agattgattt accccggttg ataatcagaa
     3121 aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaaacg ttaatatttt
     3181 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3241 cggcaaaatc ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt
     3301 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3361 ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag
     3421 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3481 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3541 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3601 gctacagggc gcgtaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3841 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4441 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagct atggtgcact
     4501 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     4561 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4621 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4681 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4741 cgtggtcgtg cagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4801 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4861 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     4921 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     4981 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     5041 ctcagggtca atgccagccg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     5101 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     5161 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     5221 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     5281 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     5341 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5401 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5461 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5521 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5581 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5641 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5701 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5761 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5821 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5881 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5941 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     6001 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     6061 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     6121 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6181 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6241 cccagttg
//
