LOCUS       sequence-448443-        5537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phage GIL01 protein gp1 (A11V) mutant with a single nucleotide 
            substitution (C32T).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5537)
  TITLE     Autoregulation ensures vertical transmission of the linear prophage 
            GIL01 in the mosquito pathogen Bacillus thuringiensis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5537)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET29b(+)_gp1(A11V)
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..27
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             307..324
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(377..395)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      391..438
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      475..930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(562..581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     772..793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1022..1837)
                     /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(1022..1837)
                     /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     1745..1764
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1829..1848
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1959..2547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2448..2467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2701..2718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2733..2872
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2798..2817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2958..2980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2974..3165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3974..5056)
                     /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     5018..5037
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5057..5134)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5340..5359
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5447..5466
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5447..5465
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5466..5490
                     /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             5505..5527
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 agaggatcgc atcaccatca ccatcacgga tccatgagta aactactgac tgcgcaagag
       61 gttgtagact tattgcgtgt tcataaaaag actatccata gaatgataca ttctggaaag
      121 ttggacgctt caaaagtagc aaacaaattc ctcattaaag aggaagatgc aaaaggacta
      181 ttggaaaata aaaataaact ggacaattaa tagcttgacc cattcgttcg ccattgtaag
      241 aataccacgc tcctgattta tcaacgattc tattacctac aatttcttgt ccttgtttaa
      301 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
      361 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      421 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
      481 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      541 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      601 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      661 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      721 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      781 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      841 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      901 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
      961 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     1021 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     1081 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     1141 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     1201 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1261 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1321 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     1381 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     1441 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     1501 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     1561 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     1621 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     1681 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     1741 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     1801 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     1861 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2101 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2581 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2641 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2701 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     2761 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     2821 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     2881 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2941 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3001 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3061 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     3121 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     3181 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     3241 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     3301 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     3361 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     3421 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     3481 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     3541 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     3601 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     3661 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     3721 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     3781 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     3841 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     3901 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     3961 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4021 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4081 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4141 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4201 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     4261 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4321 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4381 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4441 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     4501 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     4561 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     4621 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     4681 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     4741 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     4801 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     4861 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     4921 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     4981 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5041 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5101 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5161 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5221 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5281 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     5341 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     5401 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact
     5461 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     5521 aaggagatat acatatg
//
