LOCUS       sequence-6891-ex        7674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7674)
  TITLE     Collier lab plasmids
REFERENCE   2  (bases 1 to 7674)
  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"
COMMENT     Sequence Label: pET-22b PA WT
FEATURES             Location/Qualifiers
     source          1..7674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     4..69
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2317..2334
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2387..2405)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2401..2448
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2485..2940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2572..2591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2782..2803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2967..3071
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3072..3140,3141..3932)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3290..3309)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4103..4691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4592..4611
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4845..4862
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4877..5019
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4945..4964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5105..5127
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5121..5312)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6121..7203)
                     /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     7165..7184
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7204..7281)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7487..7506
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7590..7609
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7590..7608
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7609..7633
                     /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             7648..7670
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 catatgaaat acctgctgcc gaccgctgct gctggtctgc tgctcctcgc tgcccagccg
       61 gcgatggcca tggatatcgg aattaattcg gatccgatgg aagttaaaca ggagaaccgg
      121 ttattaaatg aatcagaatc aagttcccag gggttactag gatactattt tagtgatttg
      181 aattttcaag cacccatggt ggttacctct tctactacag gggatttatc tattcctagt
      241 tctgagttag aaaatattcc atcggaaaac caatattttc aatctgctat ttggtcagga
      301 tttatcaaag ttaagaagag tgatgaatat acatttgcta cttccgctga taatcatgta
      361 acaatgtggg tagatgacca agaagtgatt aataaagctt ctaattctaa caaaatcaga
      421 ttagaaaaag gaagattata tcaaataaaa attcaatatc aacgagaaaa tcctactgaa
      481 aaaggattgg atttcaagtt gtactggacc gattctcaaa ataaaaaaga agtgatttct
      541 agtgataact tacaattgcc agaattaaaa caaaaatctt cgaactcaag aaaaaagcga
      601 agtacaagtg ctggacctac ggttccagac cgtgacaatg atggaatccc tgattcatta
      661 gaggtagaag gatatacggt tgatgtcaaa aataaaagaa cttttctttc accatggatt
      721 tctaatattc atgaaaagaa aggattaacc aaatataaat catctcctga aaaatggagc
      781 acggcttctg atccgtacag tgatttcgaa aaggttacag gacggattga taagaatgta
      841 tcaccagagg caagacaccc ccttgtggca gcttatccga ttgtacatgt agatatggag
      901 aatattattc tctcaaaaaa tgaggatcaa tccacacaga atactgatag tgaaacgaga
      961 acaataagta aaaatacttc tacaagtagg acacatacta gtgaagtaca tggaaatgca
     1021 gaagtgcatg cgtcgttctt tgatattggt gggagtgtat ctgcaggatt tagtaattcg
     1081 aattcaagta cggtcgcaat tgatcattca ctatctctag caggggaaag aacttgggct
     1141 gaaacaatgg gtttaaatac cgctgataca gcaagattaa atgccaatat tagatatgta
     1201 aatactggga cggctccaat ctacaacgtg ttaccaacga cttcgttagt gttaggaaaa
     1261 aatcaaacac tcgcgacaat taaagctaag gaaaaccaat taagtcaaat acttgcacct
     1321 aataattatt atccttctaa aaacttggcg ccaatcgcat taaatgcaca agacgatttc
     1381 agttctactc caattacaat gaattacaat caatttcttg agttagaaaa aacgaaacaa
     1441 ttaagattag atacggatca agtatatggg aatatagcaa catacaattt tgaaaatgga
     1501 agagtgaggg tggatacagg ctcgaactgg agtgaagtgt taccgcaaat tcaagaaaca
     1561 actgcacgta tcatttttaa tggaaaagat ttaaatctgg tagaaaggcg gatagcggcg
     1621 gttaatccta gtgatccatt agaaacgact aaaccggata tgacattaaa agaagccctt
     1681 aaaatagcat ttggatttaa cgaaccgaat ggaaacttac aatatcaagg gaaagacata
     1741 accgaatttg attttaattt cgatcaacaa acatctcaaa atatcaagaa tcagttagcg
     1801 gaattaaacg caactaacat atatactgta ttagataaaa tcaaattaaa tgcaaaaatg
     1861 aatattttaa taagagataa acgttttcat tatgatagaa ataacatagc agttggggcg
     1921 gatgagtcag tagttaagga ggctcataga gaagtaatta attcgtcaac agagggatta
     1981 ttgttaaata ttgataagga tataagaaaa atattatcag gttatattgt agaaattgaa
     2041 gatactgaag ggcttaaaga agttataaat gacagatatg atatgttgaa tatttctagt
     2101 ttacggcaag atggaaaaac atttatagat tttaaaaaat ataatgataa attaccgtta
     2161 tatataagta atcccaatta taaggtaaat gtatatgctg ttactaaaga aaacactatt
     2221 attaatccta gtgagaatgg ggatactagt accaacggga tcaagaaaat tttaatcttt
     2281 tctaaaaaag gctatgagat aggataatga ctcgagcacc accaccacca ccactgagat
     2341 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2401 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     2461 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2521 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2581 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2641 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2701 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2761 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2821 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2881 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2941 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3001 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3061 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3121 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3181 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3241 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3301 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3361 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3421 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3481 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3541 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3601 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3661 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3721 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3781 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3841 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3901 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3961 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4021 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4081 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4141 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4201 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4261 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4321 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4381 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4441 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4501 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4561 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4621 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4681 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4741 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4801 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4861 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4921 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4981 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     5041 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5101 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     5161 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     5221 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     5281 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     5341 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     5401 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     5461 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     5521 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     5581 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     5641 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     5701 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     5761 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     5821 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     5881 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     5941 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     6001 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     6061 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     6121 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6181 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     6241 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     6301 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     6361 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     6421 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     6481 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     6541 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     6601 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     6661 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     6721 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     6781 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     6841 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     6901 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     6961 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     7021 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     7081 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     7141 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     7201 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     7261 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     7321 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     7381 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     7441 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     7501 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     7561 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     7621 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tata
//
