LOCUS       sequence-432442-        4985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid that expresses LacI, and pIII under the Lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4985)
  AUTHORS   Gretton D, Wang B, Edison R, Foner L, Berlips J, Vogel T, Kysel M, 
            Chen W, Sage-Ling F, Hauwe LV, Wooster S, Weinstein-Raun B, 
            DeBenedictis EA, Liu AB, Chory E, Cui H, Li X, Dong J, Fabrega A, 
            Dennison C, Don O, Ye CT, Uberoy K, Rivest RL, Gao M, Yu Y, Baum C, 
            Damgard I, Yao AC, Esvelt KM
  TITLE     Random adversarial threshold search enables automated DNA screening
  JOURNAL   bioRxiv 2024.03.20.585782
REFERENCE   2  (bases 1 to 4985)
  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: "bioRxiv 
            2024.03.20.585782"
COMMENT     Sequence Label: LacI-pLacI-pIII pm
FEATURES             Location/Qualifiers
     source          1..4985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      639..1094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(726..745)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     936..957
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1105..1207
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(1208..1276,1277..2068)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAVITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1426..1445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2069..2260
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      2284..2311
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2323..2910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2403..2422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2932..2961)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        3003..3080
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3081..4163
                     /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     complement(3100..3119)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    4222..4243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4258..4263,4264..4281,4282..4288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4296..4312
                     /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             join(4349..4985,1..638)
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKSHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDLFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
ORIGIN
        1 atgaggattt atttgtttgt gaatatcaag gccaatcgtc tgacctgcct caacctcctg
       61 tcaatgctgg cggcggctct ggtggtggtt ctggtggcgg ctctgagggt ggtggctctg
      121 agggtggcgg ttctgagggt ggcggctctg agggaggcgg ttccggtggt ggctctggtt
      181 ccggtgattt tgattatgaa aagatggcaa acgctaataa gggggctatg accgaaaatg
      241 ccgatgaaaa cgcgctacag tctgacgcta aaggcaaact tgattctgtc gctactgatt
      301 acggtgctgc tatcgatggt ttcattggtg acgtttccgg ccttgctaat ggtaatggtg
      361 ctactggtga ttttgctggc tctaattccc aaatggctca agtcggtgac ggtgataatt
      421 cacctttaat gaataatttc cgtcaatatt taccttccct ccctcaatcg gttgaatgtc
      481 gcccttttgt ctttggcgct ggtaaaccat atgaattttc tattgattgt gacaaaataa
      541 acttattccg tggtgtcttt gcgtttcttt tatatgttgc cacctttatg tatgtatttt
      601 ctacgtttgc taacatactg cgtaataagg agtcttaaac gcgccctgta gcggcgcatt
      661 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      721 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      781 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      841 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      901 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      961 aacactcaaa cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     1021 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1081 aacgtttaca atttgctttt gacttgatcg gcacgtaaga ggttccaact ttcaccataa
     1141 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     1201 agctaaaatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1261 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1321 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1381 acgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1441 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1501 tgacttggtt gagtactcac cagtcacaga aaagcatctc acggatggca tgacagtaag
     1561 agaattatgc agtgctgtca taaccatgag tgataacact gcggccaact tacttctggc
     1621 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1681 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1741 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1801 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggatcact
     1861 tctgcgctcg gccctcccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1921 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gcatcgtagt
     1981 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2041 aggtgcctca ctgattaagc attggtaagt gaccaaacag gaaaaaaccg cccttaacat
     2101 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     2161 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     2221 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga tgagggccca aatgtaatca
     2281 cctggctcac cttcgggtgg gcctttctgc gttgctggcg tttttccata ggctccgccc
     2341 ccctgacgag catcacaaaa atcgatgctc aagtcagagg tggcgaaacc cgacaggact
     2401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2581 cgaacccccc gttcagcccg accgctgcgc cttatcctgt aactatcgtc ttgagtccaa
     2641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2761 aagaacagta tttggtatct gcgctctgct gaagccagtt acctcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgattttcta ccgaagaaag
     2941 acccacccgt gaaggtgagc cagtgagttg attgcagtcc agttacgctg gagtcaagac
     3001 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     3061 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     3121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     3181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     3241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     3301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     3361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     3421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     3481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     3541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     3601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     3661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     3721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     3781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     3841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg
     3901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     3961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     4021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     4081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4141 cccgactgga aagcgggcag tgagcggtac ccgataaaag cggcttcctg acaggaggcc
     4201 gttttgtttt gcagcccacc ttaatgtgag ttagctcact cattaggcac cccaggcttt
     4261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4321 acctgcaggt gcagtaagga ggaaaaaagt gaaaaaatta ttattcgcaa ttcctttagt
     4381 tgttcctttc tattctcact ccgctgaaac tgttgaaagt tgtttagcaa aatcccatac
     4441 agaaaattca tttactaacg tctggaaaga cgacaaaact ttagatcgtt acgctaacta
     4501 tgagggctgt ctgtggaatg ctacaggcgt tgtagtttgt actggtgacg aaactcagtg
     4561 ttacggtaca tgggttccta ttgggcttgc tatccctgaa aatgagggtg gtggctctga
     4621 gggtggcggt tctgagggtg gcggttctga gggtggcggt actaaacctc ctgagtacgg
     4681 tgatacacct attccgggct atacttatat caaccctctc gacggcactt atccgcctgg
     4741 tactgagcaa aaccccgcta atcctaatcc ttctcttgag gagtctcagc ctcttaatac
     4801 tttcatgttt cagaataata ggttccgaaa taggcagggg gcattaactg tttatacggg
     4861 cactgttact caaggcactg accccgttaa aacttattac cagtacactc ctgtatcatc
     4921 aaaagccatg tatgacgctt actggaacgg taaattcaga gactgcgctt tccattctgg
     4981 cttta
//
