LOCUS       sequence-432443-        4941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid that expresses LacI, and pII under the Lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  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 4941)
  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-pII pm
FEATURES             Location/Qualifiers
     source          1..4941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    837..853
                     /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(861..867,868..885,886..891))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    906..927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(986..2068)
                     /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     2030..2049
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2069..2146)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      2188..2217
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2239..2826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2727..2746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2838..2865)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2889..3080)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3081..3872,3873..3941))
                     /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     3704..3723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3942..4044)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(4055..4510)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4192..4213)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4404..4423
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(4511..4941,1..802))
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGINLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVIWNETSRHRTLVAYLKHVELQHYIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
ORIGIN
        1 agcggattgc atcaaaaaga ttaagaggaa gcccgaaaga cttcaaatat cgcgttttaa
       61 ttcgagcttc aaagcgaacc agaccggaag caaactccaa caggtcagga ttagagagta
      121 cctttaattg ctccttttga taagaggtca tttttgcgga tggcttagag cttaattgct
      181 gaatataatg ctgtagctca acatgtttta aatatgcaac taaagtacgg tgtctggaag
      241 tttcattcca tataacagtt gattcccaat tctgcgaacg agtagattta gtttgaccat
      301 tagatacatt tcgcaaatgg tcaataacct gtttagctat attttcattt ggggcgcgag
      361 ctgaaaaggt ggcatcaatt ctactaatag tagtagcatt aacatccaat aaatcataca
      421 ggcaaggcaa agaattagca aaattaagca ataaagcctc agagcataaa gctaaatcgg
      481 ttgtaccaaa aacattatga ccctgtaata cttttgcggg agaagccttt atttcaacgc
      541 aaggataaaa atttttagaa ccctcatata ttttaaatgc aatgcctgag taatgtgtag
      601 gtaaagattc aaaagggtga gaaaggccgg agacagtcaa atcaccatca atatgatatt
      661 caaccgttct agctgataaa ttaatgccgg agagggtagc tatttttgag agatctacaa
      721 aggctatcag gtcattgcct gagagtctgg agcaaacaag agaatcgatg aacggtaatc
      781 gtaaaactag catgtcaatc atttttcctc cttactgcac ctgcaggtgt gtgaaattgt
      841 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      901 gcctaatgag tgagctaact cacattaagg tgggctgcaa aacaaaacgg cctcctgtca
      961 ggaagccgct tttatcgggt accgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1021 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     1081 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     1141 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     1201 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     1261 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     1321 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     1381 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     1441 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     1501 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     1561 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     1621 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     1681 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     1741 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     1801 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     1861 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     1921 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     1981 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     2041 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     2101 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtcgggt cttgactcca
     2161 gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggtct ttcttcggta
     2221 gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2281 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2341 tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2521 cgatagttac aggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2761 tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta
     2821 tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacatttg
     2881 ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     2941 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     3001 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     3061 gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatgcgg
     3181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3241 ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag tgatcctgca
     3301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag
     3541 ttggccgcag tgttatcact catggttatg acagcactgc ataattctct tactgtcatg
     3601 ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa actctcaagg
     3781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3901 aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc cttagctcct
     3961 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     4021 ttggaacctc ttacgtgccg atcaagtcaa aagcaaattg taaacgttaa tattttgtta
     4081 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4141 aaaatccctt ataaatcaaa agaatagccc gagataggtt tgagtgttgt tccagtttgg
     4201 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4261 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     4321 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4381 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4441 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4501 cagggcgcgt ttatgcgatt ttaagaactg gctcattata ccagtcagga cgttgggaag
     4561 aaaaatctac gttaataaaa cgaactaacg gaacaacatt attacaggta gaaagattca
     4621 tcagttgaga tttaggaata ccacattcaa ctaatgcaga tacataacgc caaaaggaat
     4681 tacgaggcat agtaagagca acactatcat aaccctcgtt taccagacga cgataaaaac
     4741 caaaatagcg agaggctttt gcaaaagaag ttttgccaga gggggtaata gtaaaatgtt
     4801 tagactggat agcgtccaat actgcggaat cgtcataaat attcattgaa tccccctcaa
     4861 atgctttaaa cagttcagaa aacgagaatg accataaatc aaaaatcagg tctttaccct
     4921 gactattata gtcagaagca a
//
