LOCUS       sequence-385961-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses phage Av05 NTase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  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 6463)
  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-Av05  NTase
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..167
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     255..274
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        255..273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    274..298
                     /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             313..335
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             342..455
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             474..494
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1551..1568
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1575..1595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(1648..1666)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1662..1709
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1746..2201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1833..1852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2043..2064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2293..3108)
                     /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(2293..3108)
                     /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     3016..3035
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3100..3119
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3230..3818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3719..3738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3972..3989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4004..4143
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4069..4088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4229..4251
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4245..4436)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5245..6327)
                     /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     6289..6308
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6328..6405)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
       61 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
      121 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
      181 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct
      241 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
      301 tctagaaata attttgttta actttaagaa ggagatatac aatggatgaa aaaaccaccg
      361 gatggcgtgg ggggcatgtg gtagaaggat tagcaggaga gttagaacaa ttacgtgctc
      421 gtttagaaca ccacccacag ggacagcgcg agcccggtgg gagtgggtca ggagaaaacc
      481 tctacttcca aggtaagaaa ttggaattga tgcgttgcta ctctgggagc atcgcgtatg
      541 gcacaaatct tcccacctcc gatgttgaca ttcgtggaat ctactgcgca gagcccaagt
      601 ccatcttgac gccattcttt gtcgatcgtg agatgaccct gattgatgaa gaagatggaa
      661 agctgtatga attgaccaat tatatgaagc tgtttgtcga tatgaaccct aatattattg
      721 agttaatgtt cgttgatcgt ggtgacattt tgcaaacctc agacgtctac gaatacttac
      781 gtgaacaagc acccaaactg ttgtctagca aggtagcttt tacattctca ggatacgcca
      841 tggcccaatt aaaacgtatt aaaggacaca ataaatggat caataaccct cagcccaaag
      901 aacagccaac tcaaaaggag ttcatgcgct taacccatag ttggatggac gataatctga
      961 ccaagcacaa agatttccag aagatgttag atgatcttaa taaattgtgt gtgttcctgc
     1021 ccatgggaga caacatctac ggtgtaattg ggatccacga caacgccggc ttatttaact
     1081 ctgacggtag cattcgcaaa gtcccttatg aacaattctc agatgaggat aaaaaacgta
     1141 agcccatcct gatcgtaaag tatttggcag atgatcacaa gcaggccaag gagaagcaca
     1201 aaaactactg ggcgtgggtt gaaaatcgca acccagttcg ccatgaattg gaagaaaaat
     1261 atggctatga ctgtaaacac gctcttcacc tggttcgtct gatgcgtatg gccgaagaaa
     1321 tcttggcaga aggaaaggta ttggttaaac gtcctgatgc acaggaactg ctggcaatcc
     1381 gtaatggcgc atggtctctt gaaaagttac tggcctgggc tgaggagaaa gacgattata
     1441 ttcgcaatac tttatacaaa aacactcatt tgcccaagac cacagattta aactttgctg
     1501 cggacgtact tatgacggct cagcagatgt gttgggacaa aatgaacgac caccaccacc
     1561 atcaccatta gtgacaccac caccaccacc accactgaga tccggctgct aacaaagccc
     1621 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1681 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
     1741 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1801 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1861 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1921 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1981 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2041 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2101 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2161 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2281 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     2341 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     2401 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2461 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2521 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     2581 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2641 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     2701 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2761 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     2821 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2881 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2941 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     3001 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     3061 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     3121 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     3181 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3301 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3361 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3421 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3481 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3541 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3601 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3661 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3781 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3841 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3901 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3961 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     4021 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     4081 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4141 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4201 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4261 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4321 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4381 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     4441 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     4501 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     4561 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     4621 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     4681 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     4741 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     4801 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     4861 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     4921 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     4981 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     5041 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     5101 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5161 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5221 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5281 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5341 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5401 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5461 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5521 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5581 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5641 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5701 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5761 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5821 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     5881 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     5941 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     6001 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     6061 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     6121 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     6181 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     6241 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6301 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6361 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6421 ctcccttatg cgactcctgc attaggaagc agcccagtag tag
//
