LOCUS       sequence-486013-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAD-sd8-GIII Cassette; AP Pos Circuit for AraC phage-assisted 
            evolution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6855)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 6855)
  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: PACE_AP_Pos_pBADProm
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..951)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      999..1221
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_feature    1834..1853
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        1860..2144
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2067..2086
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2211..3485
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     CDS             3485..4564
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     CDS             4613..5593
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
                     SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
                     SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
                     PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
                     HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
                     AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
     terminator      complement(5687..5733)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(5784..6575,6576..6644))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6407..6426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6645..6749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6817..6835
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
     1741 aagactgggc ctttcgtttt gctgaggaga cttagggacc catgccttta attaaggagt
     1801 tttgcaggtg ccttggaaca cctgcttttc gctgaattcg cggccgcttc tagactagaa
     1861 agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt
     1921 ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa
     1981 agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga
     2041 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga
     2101 tcctacctga cgctttttat cgcaactctc tactgtttct ccatgaccga attcaaaaga
     2161 tcttttaaga aggagatata catcctgcag gtgcagtaaa ggaaaaaaaa atgaaaaaat
     2221 tattattcgc aattccttta gttgttcctt tctattctca ctccgctgaa actgttgaaa
     2281 gttgtttagc aaaaccccat acagaaaatt catttactaa cgtctggaaa gacgacaaaa
     2341 ctttagatcg ttacgctaac tatgagggct gtctgtggaa tgctacaggc gttgtagttt
     2401 gtactggtga cgaaactcag tgttacggta catgggttcc tattgggctt gctatccctg
     2461 aaaatgaggg tggtggctct gagggtggcg gttctgaggg tggcggttct gagggtggcg
     2521 gtactaaacc tcctgagtac ggtgatacac ctattccggg ctatacttat atcaaccctc
     2581 tcgacggcac ttatccgcct ggtactgagc aaaaccccgc taatcctaat ccttctcttg
     2641 aggagtctca gcctcttaat actttcatgt ttcagaataa taggttccga aataggcagg
     2701 gggcattaac tgtttatacg ggcactgtta ctcaaggcac tgaccccgtt aaaacttatt
     2761 accagtacac tcctgtatca tcaaaagcca tgtatgacgc ttactggaac ggtaaattca
     2821 gagactgcgc tttccattct ggctttaatg aggatccatt cgtttgtgaa tatcaaggcc
     2881 aatcgtctga cctgcctcaa cctcctgtca atgctggcgg cggctctggt ggtggttctg
     2941 gtggcggctc tgagggtggt ggctctgagg gtggcggttc tgagggtggc ggctctgagg
     3001 gaggcggttc cggtggtggc tctggttccg gtgattttga ttatgaaaag atggcaaacg
     3061 ctaataaggg ggctatgacc gaaaatgccg atgaaaacgc gctacagtct gacgctaaag
     3121 gcaaacttga ttctgtcgct actgattacg gtgctgctat cgatggtttc attggtgacg
     3181 tttccggcct tgctaatggt aatggtgcta ctggtgattt tgctggctct aattcccaaa
     3241 tggctcaagt cggtgacggt gataattcac ctttaatgaa taatttccgt caatatttac
     3301 cttccctccc tcaatcggtt gaatgtcgcc cttttgtctt tggcgctggt aaaccttacg
     3361 agttcagtat cgactgcgat aagatcaacc tgttccgcgg tgtctttgcg tttcttttat
     3421 atgttgccac ctttatgtat gtattttcta cgtttgctaa catactgcgt aataaggagt
     3481 cttaatgaaa tttggaaact ttttgcttac ataccaacct ccccaatttt cccaaacaga
     3541 ggtaatgaaa cgtttggtta aattaggtcg catctctgag gagtgtggtt ttgataccgt
     3601 atggttactg gagcatcatt tcacggagtt tggtttgctt ggtaaccctt atgtcgctgc
     3661 tgcatattta cttggcgcga ctaaaaaatt gaatgtagga actgccgcta ttgttcttcc
     3721 cacagcccat ccagtacgcc aacttgaaga tgtgaattta ttggatcaaa tgtcaaaagg
     3781 acgatttcgg tttggtattt gccgagggct ttacaacaag gactttcgcg tattcggcac
     3841 agatatgaat aacagtcgcg ccttagcgga atgctggtac gggctgataa agaatggcat
     3901 gacagaggga tatatggaag ctgataatga acatatcaag ttccataagg taaaagtaaa
     3961 ccccgcggcg tatagcagag gtggcgcacc ggtttatgtg gtggctgaat cagcttcgac
     4021 gactgagtgg gctgctcaat ttggcctacc gatgatatta agttggatta taaatactaa
     4081 cgaaaagaaa gcacaacttg agctttataa tgaagtggct caagaatatg ggcacgatat
     4141 tcataatatc gaccattgct tatcatatat aacatctgta gatcatgact caattaaagc
     4201 gaaagagatt tgccggaaat ttctggggca ttggtatgat tcttatgtga atgctacgac
     4261 tatttttgat gattcagacc aaacaagagg ttatgatttc aataaagggc agtggcgtga
     4321 ctttgtatta aaaggacata aagatactaa tcgccgtatt gattacagtt acgaaatcaa
     4381 tcccgtggga acgccgcagg aatgtattga cataattcaa aaagacattg atgctacagg
     4441 aatatcaaat atttgttgtg gatttgaagc taatggaaca gtagacgaaa ttattgcttc
     4501 catgaagctc ttccagtctg atgtcatgcc atttcttaaa gaaaaacaac gttcgctatt
     4561 atattatggc ggtggcggta gcggcggtgg cggtagcggc ggtggcggta gcaaatttgg
     4621 attgttcttc cttaacttca tcaattcaac aactgttcaa gaacagagta tagttcgcat
     4681 gcaggaaata acggagtatg ttgataagtt gaattttgaa cagattttag tgtatgaaaa
     4741 tcatttttca gataatggtg ttgtcggcgc tcctctgact gtttctggtt ttctgctcgg
     4801 tttaacagag aaaattaaaa ttggttcatt aaatcacatc attacaactc atcatcctgt
     4861 ccgcatagcg gaggaagctt gcttattgga tcagttaagt gaagggagat ttattttagg
     4921 gtttagtgat tgcgaaaaaa aagatgaaat gcattttttt aatcgcccgg ttgaatatca
     4981 acagcaacta tttgaagagt gttatgaaat cattaacgat gctttaacaa caggctattg
     5041 taatccagat aacgattttt atagcttccc taaaatatct gtaaatcccc atgcttatac
     5101 gccaggcgga cctcggaaat atgtaacagc aaccagtcat catattgttg agtgggcggc
     5161 caaaaaaggt attcctctca tctttaagtg ggatgattct aatgatgtta gatatgaata
     5221 tgctgaaaga tataaagccg ttgcggataa atatgacgtt gacctatcag agatagacca
     5281 tcagttaatg atattagtta actataacga agatagtaat aaagctaaac aagagacgcg
     5341 tgcatttatt agtgattatg ttcttgaaat gcaccctaat gaaaatttcg aaaataaact
     5401 tgaagaaata attgcagaaa acgctgtcgg aaattatacg gagtgtataa ctgcggctaa
     5461 gttggcaatt gaaaagtgtg gtgcgaaaag tgtattgctg tcctttgaac caatgaatga
     5521 tttgatgagc caaaaaaatg taatcaatat tgttgatgat aatattaaga agtaccacac
     5581 ggaatatacc taaacttaat taacggcact cctcagcaaa tataatgacc ctcttgataa
     5641 cccaagaggg cattttttaa tgcccatggc gtagctgtgc tctagaataa aacgaaaggc
     5701 ccagtctttc gactgagcct ttcgttttat ttgaccggaa atcagaattg gttaattggt
     5761 tgtaacactg gcagagcatt acgttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6781 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggccctta
     6841 ggccaaatag gccgt
//
