LOCUS       sequence-393185-        5506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expersses phage MazG-like/Atd1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5506)
  AUTHORS   Ho P, Chen Y, Biswas S, Canfield E, Abdolvahabi A, Feldman DE
  TITLE     Bacteriophage antidefense genes that neutralize TIR and STING immune
            responses.
  JOURNAL   Cell Rep. 2023 Mar 22;42(4):112305. doi: 
            10.1016/j.celrep.2023.112305.
  PUBMED    36952342
REFERENCE   2  (bases 1 to 5506)
  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: "Cell Rep.";
            date: "2023-03-22"; volume: "42(4):112305. doi"; pages: " 
            10.1016/j.celrep.2023.112305"
COMMENT     Sequence Label: pET30a- phage MazG-like/Atd1
FEATURES             Location/Qualifiers
     source          1..5506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(387..1469)
                     /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     1431..1450
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1470..1547)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1753..1772
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1860..1879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1860..1878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1879..1903
                     /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             1918..1940
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1950..1973
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2296..2314)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2310..2357
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2394..2849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2481..2500)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2691..2712
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2941..3756)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2941..3756)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3664..3683
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3748..3767
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3878..4466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4367..4386
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4620..4637
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4652..4791
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4717..4736)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4877..4899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4893..5084)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
       61 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
      121 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
      181 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
      241 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
      301 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
      361 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      421 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
      481 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
      541 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
      601 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
      661 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
      721 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
      781 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
      841 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
      901 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
      961 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     1021 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     1081 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     1141 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     1201 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     1261 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     1321 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     1381 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     1441 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     1501 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     1561 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     1621 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     1681 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     1741 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     1801 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt
     1861 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     1921 gtttaacttt aagaaggaga tatacaatgg actacaaaga cgatgacgat aaagaggaca
     1981 agccggggta tcacttggat aaaatcaaaa agcgccgttt tggtagttta ggtaagatcc
     2041 aggaggaggt agaagagttg gtcgatgcgc accgtcaagg atcgcatgtc atgattctgg
     2101 tggagatgtc ggacctgtat ggagctatgc aaggatttct tgaggagaac tatccaggct
     2161 ttaaaatgga ggatcttaag aaattttcgg gcatcacaca gcgtgccttt aaaaacggtt
     2221 accgttagca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg
     2281 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     2341 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     2401 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2461 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2521 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2581 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2641 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2701 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2761 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2821 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     2881 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     2941 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     3001 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     3061 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     3121 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     3181 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     3241 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     3301 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     3361 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     3421 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     3481 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     3541 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     3601 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     3661 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     3721 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     3781 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     3841 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3901 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3961 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4021 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4081 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4141 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4201 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4261 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4321 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4381 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4441 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4501 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4561 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4621 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4681 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4741 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4801 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4861 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4921 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4981 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     5041 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     5101 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     5161 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     5221 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     5281 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     5341 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     5401 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     5461 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagc
//
