LOCUS       sequence-432462-        4754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid that expresses LacI, and pI under the Lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4754)
  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 4754)
  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-pI pm
FEATURES             Location/Qualifiers
     source          1..4754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    743..759
                     /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)."
     promoter        complement(join(767..773,774..791,792..797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    812..833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(892..1974)
                     /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     1936..1955
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1975..2052)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      2094..2123
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2145..2732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2633..2652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2744..2771)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2795..2986)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(2987..3778,3779..3847))
                     /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     3610..3629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3848..3950)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(3961..4416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4098..4119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4310..4329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(4417..4754,1..709))
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
ORIGIN
        1 aatttaatgg tttgaaatac cgaccgtgtg ataaataagg cgttaaataa gaataaacac
       61 cggaatcata attactagaa aaagcctgtt tagtatcata tgcgttatac aaattcttac
      121 cagtataaag ccaacgctca acagtagggc ttaattgaga atcgccatat ttaacaacgc
      181 caacatgtaa tttaggcaga ggcattttcg agccagtaat aagagaatat aaagtaccga
      241 caaaaggtaa agtaattctg tccagacgac gacaataaac aacatgttca gctaatgcag
      301 aacgcgcctg tttatcaaca atagataagt cctgaacaag aaaaataata tcccatccta
      361 atttacgagc atgtagaaac caatcaataa tcggctgtct ttccttatca ttccaagaac
      421 gggtattaaa ccaagtaccg cactcatcga gaacaagcaa gccgttttta ttttcatcgt
      481 aggaatcatt accgcgccca atagcaagca aatcagatat agaaggctta tccggtattc
      541 taagaacgcg aggcgtttta gcgaacctcc cgacttgcgg gaggttttga agccttaaat
      601 caagattagt tgctattttg cacccagcta caattttatc ctgaatctta ccaacgctaa
      661 cgagcgtctt tccagagcct aatttgccag ttacaaaata aacagccatt ttcctcctta
      721 ctgcacctgc aggtgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      781 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaaggtggg
      841 ctgcaaaaca aaacggcctc ctgtcaggaa gccgctttta tcgggtaccg ctcactgccc
      901 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      961 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     1021 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     1081 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     1141 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     1201 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     1261 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     1321 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     1381 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     1441 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     1501 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     1561 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     1621 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     1681 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     1741 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     1801 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     1861 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     1921 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     1981 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     2041 ttcgatggtg tcgggtcttg actccagcgt aactggactg caatcaactc actggctcac
     2101 cttcacgggt gggtctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     2161 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2221 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     2281 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2341 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2401 cgtgtcttac cgggttggac tcaagacgat agttacagga taaggcgcag cggtcgggct
     2461 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2521 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2581 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2641 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     2701 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     2761 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     2821 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     2881 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     2941 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     3001 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3061 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     3121 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     3181 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     3241 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3301 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3361 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3421 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatgacag
     3481 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     3541 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3601 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3661 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3721 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3781 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3841 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     3901 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     3961 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4021 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     4081 taggtttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4141 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     4201 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4261 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4321 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4381 cacccgccgc gcttaatgcg ccgctacagg gcgcgtttaa ttacatttaa caatttcatt
     4441 tgaattacct tttttaatgg aaacagtaca taaatcaata tatgtgagtg aataaccttg
     4501 cttctgtaaa tcgtcgctat taattaattt tcccttagaa tccttgaaaa catagcgata
     4561 gcttagatta agacgctgag aagagtcaat agtgaattta tcaaaatcat aggtctgaga
     4621 gactaccttt ttaacctccg gcttaggttg ggttatataa ctatatgtaa atgctgatgc
     4681 aaatccaatc gcaagacaaa gaacgcgaga aaactttttc aaatatattt tagttaattt
     4741 catcttctga ccta
//
