LOCUS       sequence-432441-        3583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid that expresses LacI but no phage proteins, used as a 
            control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3583)
  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 3583)
  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 pm
FEATURES             Location/Qualifiers
     source          1..3583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     402..421
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(441..518)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      560..589
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      611..1198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1099..1118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1210..1237)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1261..1452)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1453..2244,2245..2313))
                     /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     2076..2095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2314..2416)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(2427..2882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2564..2585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2776..2795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(2941..3583,1..440))
                     /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"
ORIGIN
        1 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
       61 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
      121 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
      181 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
      241 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
      301 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
      361 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
      421 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
      481 gccataccgc gaaaggtttt gcgccattcg atggtgtcgg gtcttgactc cagcgtaact
      541 ggactgcaat caactcactg gctcaccttc acgggtgggt ctttcttcgg tagaaaatca
      601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
      721 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      901 acaggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1141 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1201 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     1261 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     1321 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     1381 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     1441 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     1561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1621 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     1681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     1921 agtgttatca ctcatggtta tgacagcact gcataattct cttactgtca tgccatccgt
     1981 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     2161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2281 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     2341 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     2401 tcttacgtgc cgatcaagtc aaaagcaaat tgtaaacgtt aatattttgt taaaattcgc
     2461 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2521 ttataaatca aaagaatagc ccgagatagg tttgagtgtt gttccagttt ggaacaagag
     2581 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2641 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     2701 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2761 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2821 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2881 gtaggtgggc tgcaaaacaa aacggcctcc tgtcaggaag ccgcttttat cgggtaccgc
     2941 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3001 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     3061 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     3121 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     3181 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     3241 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     3301 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     3361 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     3421 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     3481 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     3541 gaaaataata ctgttgatgg gtgtctggtc agagacatca aga
//
