LOCUS       sequence-236046-        3187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly CD part for LacI-MVY (Adam Meyer's 
            improved LacI repressor from https://doi.org/10.1101/285866, 
            selected for dynamic range and low cross-sensitivity)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3187)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 3187)
  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"
COMMENT     Sequence Label: C72m
FEATURES             Location/Qualifiers
     source          1..3187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..38
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(42..85)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    88..109
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             125..1207
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(144..163)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    1218..1238
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1244..1261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1415..2003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1495..1514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2174..2965,2966..3034))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2797..2816
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3035..3139)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc
       61 cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagaga ctagtgggtc
      121 tcaaatgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatatgac
      181 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
      241 agcggcgatg gtggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
      301 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
      361 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
      421 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
      481 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
      541 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
      601 tatttactcc catgaggacg gtacgcgact gggcgtggag catctggtcg cattgggtca
      661 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
      721 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
      781 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
      841 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
      901 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagatag
      961 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     1021 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     1081 gccagtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     1141 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1201 gcagtgaggt agagacctac tagtagcggc cgctgcaggc ttcctcgctc actgactcgc
     1261 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg
     1441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1921 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1981 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2041 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2101 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2161 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2221 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2281 taccatctgg ccccagtgct gcaatgatac cgcgcgaccc acgctcaccg gctccagatt
     2341 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2401 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2461 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2521 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2581 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2641 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2701 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2761 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2821 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2881 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2941 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3001 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3061 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3121 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3181 ttattat
//
