LOCUS       sequence-83235-e        6460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derivative of pT7-agrBD-I with a point mutation in the agrD gene to 
            change AIP coding region from type-I to type-IV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6460)
  AUTHORS   Marchand N, Collins CH
  TITLE     Peptide-based communication system enables Escherichia coli to 
            Bacillus megaterium interspecies signaling.
  JOURNAL   Biotechnol Bioeng. 2013 Nov;110(11):3003-12. doi: 10.1002/bit.24975.
            Epub 2013 Jul 5.
  PUBMED    23775238
REFERENCE   2  (bases 1 to 6460)
  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: "10.1002/bit.24975";
            journalName: "Biotechnol Bioeng"; date: "2013-11"; volume: "110"; 
            issue: "11"; pages: "3003-12"
COMMENT     Sequence Label: pT7-agrBD-IV
FEATURES             Location/Qualifiers
     source          1..6460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     CDS             1040..1081
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1091..1108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1165..1183)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1179..1226
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1401..1406,1407..1423,1424..1429))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1460..1478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1546..1650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1651..1719,1720..2511)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1869..1888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2682..3270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3171..3190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3424..3441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3456..3595
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3521..3540)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3681..3703
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3697..3888)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5200..6282)
                     /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     6244..6263
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6283..6360)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggaatt caggagccct
      301 tcaccatgaa ttattttgat aataaaattg accagtttgc cacgtatctt caaaagagaa
      361 ataacttaga tcatattcaa tttttgcaag tacgattagg gatgcaggtc ttagctaaaa
      421 atataggtaa attaattgtt atgtatacta ttgcctatat tttaaacatt tttctgttta
      481 cgttaattac gaatttaaca ttttatttaa taagaagaca tgcacatggt gcacatgcac
      541 cttcttcttt ttggtgttat gtagaaagta ttatactatt tatactttta cctttagtaa
      601 tagtaaattt tcatattaac tttttaatta tgattatttt aacagttatt tctttaggtg
      661 taatctcagt atatgctcct gcagcaacta aaaagaagcc cattcctgtg cgacttatta
      721 aacgaaaaaa atattatgcg attattgtta gtttaaccct tttcattatc acacttatca
      781 tcaaagagcc atttgcccaa ttcattcaat taggcatcat aatagaagct attacattat
      841 tacctatttt ctttattaag gaggacttaa aatgaataca ttatttaact tattttttga
      901 ttttattact gggattttaa aaaacattgg taacatcgca gcttatagta cttgttactt
      961 cataatggat gaagttgaag taccaaaaga attaacacaa ttacacgaat aaaagggcga
     1021 gctcaattcg aagcttgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     1081 gcgtaccggt catcatcacc atcaccattg agtttgatcc ggctgctaac aaagcccgaa
     1141 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1201 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa
     1261 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     1321 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     1381 taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     1441 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1861 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     1921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2161 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2821 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3421 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3481 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     3541 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3721 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3781 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3841 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     3901 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     3961 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     4021 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     4081 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     4141 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     4201 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     4261 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     4321 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     4381 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     4441 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     4501 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     4561 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     4621 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt
     4681 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     4741 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     4801 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     4861 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4921 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4981 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     5041 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     5101 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     5161 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5221 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5281 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     5341 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     5401 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5461 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5521 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5581 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5641 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5701 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5761 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5821 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5881 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5941 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     6001 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     6061 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     6121 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     6181 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     6241 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     6301 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     6361 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6421 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
//
