LOCUS       sequence-432440-        4976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid that expresses LacI, and pIV under the Lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4976)
  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 4976)
  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-pIV pm
FEATURES             Location/Qualifiers
     source          1..4976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    854..870
                     /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(878..884,885..902,903..908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    923..944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(1003..2085)
                     /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     2047..2066
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2086..2163)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      2205..2234
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2256..2843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2744..2763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2855..2882)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2906..3097)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3098..3889,3890..3958))
                     /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     3721..3740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3959..4061)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(4072..4527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4209..4230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4421..4440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(4515..4976,1..819))
                     /codon_start=1
                     /gene="IV"
                     /product="pIV"
                     /label=M13 gene IV
                     /note="phage assembly"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQSGES
                     VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSNPSIIQKYNPNNQDYIDE
                     LPSSDNQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
                     LSIDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
                     KGSISVGQNVPFITGRVTGESANVNNPFQTIERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
ORIGIN
        1 tggcacagac aatatttttg aatggctatt agtctttaat gcgcgaactg atagccctaa
       61 aacatcgcca ttaaaaatac cgaacgaacc accagcagaa gataaaacag aggtgaggcg
      121 gtcagtatta acaccgcctg caacagtgcc acgctgagag ccagcagcaa atgaaaaatc
      181 taaagcatca ccttgctgaa cctcaaatat caaaccctca atcaatatct ggtcagttgg
      241 caaatcaaca gttgaaagga attgaggaag gttatctaaa atatctttag gagcactaac
      301 aactaataga ttagagccgt caatagataa tacatttgag gatttagaag tattagactt
      361 tacaaacaat tcgacaactc gtattaaatc ctttgcccga acgttattaa ttttaaaagt
      421 ttgagtaaca ttatcatttt gcggaacaaa gaaaccacca gaaggagcgg aattatcatc
      481 atattcctga ttatcagatg atggcaattc atcaatataa tcctgattgt ttggattata
      541 cttctgaata atggaagggt tagaacctac catatcaaaa ttatttgcac gtaaaacaga
      601 aataaagaaa ttgcgtagat tttcaggttt aacgtcagat gaatatacag taacagtacc
      661 ttttacatcg ggagaaacaa taacggattc gcctgattgc tttgaatacc aagttacaaa
      721 atcgcgcaga ggcgaattat tcatttcaat tacctgagca aaagaagatg atgaaacaaa
      781 catcaagaaa acaaaattaa ttacatttaa caatttcatt tgaattacct tctgcacctg
      841 caggtgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaaggtgg gctgcaaaac
      961 aaaacggcct cctgtcagga agccgctttt atcgggtacc gctcactgcc cgctttccag
     1021 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1081 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     1141 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     1201 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     1261 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     1321 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     1381 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     1441 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     1501 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     1561 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     1621 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     1681 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     1741 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     1801 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     1861 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     1921 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     1981 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     2041 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     2101 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     2161 gtcgggtctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     2221 tgggtctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2281 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2341 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     2401 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2521 ccgggttgga ctcaagacga tagttacagg ataaggcgca gcggtcgggc tgaacggggg
     2581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2761 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     2821 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     2881 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     2941 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3001 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3061 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     3121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3181 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3241 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     3301 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3541 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatgaca gcactgcata
     3601 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     3661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3781 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     3961 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     4021 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag caaattgtaa
     4081 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4141 aataggccga aatcggcaaa atcccttata aatcaaaaga atagcccgag ataggtttga
     4201 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4261 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     4321 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4381 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4441 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4501 cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgagcac gtataacgtg
     4561 ctttcctcgt tagaatcaga gcgggagcta aacaggaggc cgattaaagg gattttagac
     4621 aggaacggta cgccagaatc ctgagaagtg tttttataat cagtgaggcc accgagtaaa
     4681 agagtctgtc catcacgcaa attaaccgtt gtagcaatac ttctttgatt agtaataaca
     4741 tcacttgcct gagtagaaga actcaaacta tcggccttgc tggtaatatc cagaacaata
     4801 ttaccgccag ccattgcaac aggaaaaacg ctcatggaaa tacctacatt ttgacgctca
     4861 atcgtctgaa atggattatt tacattggca gattcaccag tcacacgacc agtaataaaa
     4921 gggacattct ggccaacaga gatagaaccc ttctgacctg aaagcgtaag aatacg
//
