LOCUS       sequence-169620-        7095 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes araC and LambdaCI. To be used with pOP549. Encrypts pOP549. 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7095)
  TITLE     Tim Lu lab plasmids
REFERENCE   2  (bases 1 to 7095)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pOP523
FEATURES             Location/Qualifiers
     source          1..7095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     135..157
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..166
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     176..195
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        176..194
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        join(221..226,227..243,244..250)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    258..274
                     /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)."
     CDS             315..1193
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      1199..1285
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1375..1391
                     /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)."
     CDS             1593..1622
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    1800..1819
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    1833..1849
                     /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)."
     CDS             1914..2627
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     primer_bind     complement(2661..2677)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2662..2678)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2707..2727)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2707..2725)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2746..2762)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2746..2762)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2770..2786
                     /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(2794..2800,2801..2818,2819..2824))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2839..2860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     3521..3540
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3607..3709)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3826..3844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5102..5873
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5874..6536
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
       61 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      121 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
      181 cgactcacta tagggcgaat tggagctctg aaatgagctg ttgacaatta atcatccggc
      241 tcgtataatg tgtggaattg tgagcggata acaatttcac accgccttcg gcgaagctag
      301 ggacgagagc tagcatggct gaagcgcaaa atgatcccct gctgccggga tactcgttta
      361 acgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc gattttttta
      421 tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc ggtcaggggg
      481 tggtgaaaaa tcagggacga gaatttgtct gccgaccggg tgatattttg ctgttcccgc
      541 caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat caccagtggg
      601 tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca atatttgcca
      661 atacgggttt ctttcgcccg gatgaagcgc accagccgca tttcagcgac ctgtttgggc
      721 aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg ataaatctgc
      781 ttgagcaatt gttactgcgg cgcatggaag cgattaacga gtcgctccat ccgccgatgg
      841 ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac agcaattttg
      901 atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca catcttttcc
      961 gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc agccaggcga
     1021 agctgctttt gagcactacc cggatgccta tcgccaccgt cggtcgcaat gttggttttg
     1081 acgatcaact ctatttctcg cgagtattta aaaaatgcac cggggccagc ccgagcgagt
     1141 ttcgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca taaggcatca
     1201 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1261 gaacgctctc ctgagtagga caaatccgcc gccctagagc ccggatgatc ctgacagctc
     1321 caccgcggtg gcggccgctc tagaactagt ggttgaccag acaaaccacg acaattgtga
     1381 gcggataaca atttcacaca gggccctcgg acaccgagga gaatgtcaag aggcgaacac
     1441 acaacgtctt ggagcgccag aggaggaacg agctaaaacg gagctttttt gccctgcgtg
     1501 accagatccc ggagttggaa aacaatgaaa aggcccccaa ggtagttatc cttaaaaaag
     1561 ccacagcata catcctgtcc gtccaagcag aggagcaaaa gctcatttct gaagaggact
     1621 tgttgcggaa acgacgagaa cagttgaaac acaaacttga acagctacgg aactcttgtg
     1681 cgtaaggaaa agtaaggaaa acgattcctt ctaacagaaa tgtcctgagc aatcacctat
     1741 gaactgtcga ctcgagcata gcatttttat ccataagatt agcggatcct aagctttaca
     1801 attgtgagcg ctcacaatta tgatagattc aattgtgagc ggataacaat ttcacacagc
     1861 aaaaaaagca aaaagcattt accataggtc ccaatataag gagagggtgt acaatgagca
     1921 caaaaaagaa accattaaca caagagcagc ttgaggacgc acgtcgcctt aaagcaattt
     1981 atgaaaaaaa gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac aagatgggga
     2041 tggggcagtc aggcgttggt gctttattta atggcatcaa tgcattaaat gcttataacg
     2101 ccgcattgct tgcaaaaatt ctcaaagtta gcgttgaaga atttagccct tcaatcgcca
     2161 gagaaatcta cgagatgtat gaagcggtta gtatgcagcc gtcacttaga agtgagtatg
     2221 agtaccctgt tttttctcat gttcaggcag ggatgttctc acctgagctt agaaccttta
     2281 ccaaaggtga tgcggagaga tgggtaagca caaccaaaaa agccagtgat tctgcattct
     2341 ggcttgaggt tgaaggtaat tccatgaccg caccaacagg ctccaagcca agctttcctg
     2401 acggaatgtt aattctcgtt gaccctgagc aggctgttga gccaggtgat ttctgcatag
     2461 ccagacttgg gggtgatgag tttaccttca agaaactgat cagggatagc ggtcaggtgt
     2521 ttttacaacc actaaaccca cagtacccaa tgatcccatg caatgagagt tgttccgttg
     2581 tggggaaagt tatcgctagt cagtggcctg aagagacgtt tggctgaccc cgggctgcag
     2641 gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccagctt
     2701 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     2761 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2821 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2881 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2941 gagaggcggt ttgcgtattg ggcgcatgca taaaaactgt tgtaattcat taagcattct
     3001 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     3061 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     3121 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     3181 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     3241 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     3301 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     3361 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     3421 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     3481 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     3541 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     3601 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     3661 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     3721 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     3781 tgcgaagtga tcttccgtca caggtattta ttcgaagacg aaagggcctc gtgatacgcc
     3841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3901 ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc gctggacttc
     3961 ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg ccttggcctg
     4021 catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct cccgaaggtc
     4081 tcgggccgtc tcttgggctt gatcggcctt cttgcgcatc tcacgcgctc ctgcggcggc
     4141 ctgtagggca ggctcatacc cctgccgaac cgcttttgtc agccggtcgg ccacggcttc
     4201 cggcgtctca acgcgctttg agattcccag cttttcggcc aatccctgcg gtgcataggc
     4261 gcgtggctcg accgcttgcg ggctgatggt gacgtggccc actggtggcc gctccagggc
     4321 ctcgtagaac gcctgaatgc gcgtgtgacg tgccttgctg ccctcgatgc cccgttgcag
     4381 ccctagatcg gccacagcgg ccgcaaacgt ggtctggtcg cgggtcatct gcgctttgtt
     4441 gccgatgaac tccttggccg acagcctgcc gtcctgcgtc agcggcacca cgaacgcggt
     4501 catgtgcggg ctggtttcgt cacggtggat gctggccgtc acgatgcgat ccgccccgta
     4561 cttgtccgcc agccacttgt gcgccttctc gaagaacgcc gcctgctgtt cttggctggc
     4621 cgacttccac cattccgggc tggccgtcat gacgtactcg accgccaaca cagcgtcctt
     4681 gcgccgcttc tctggcagca actcgcgcag tcggcccatc gcttcatcgg tgctgctggc
     4741 cgcccagtgc tcgttctctg gcgtcctgct ggcgtcagcg ttgggcgtct cgcgctcgcg
     4801 gtaggcgtgc ttgagactgg ccgccacgtt gcccattttc gccagcttct tgcatcgcat
     4861 gatcgcgtat gccgccatgc ctgcccctcc cttttggtgt ccaaccggct cgacgggggc
     4921 agcgcaaggc ggtgcctccg gcgggccact caatgcttga gtatactcac tagactttgc
     4981 ttcgcaaagt cgtgaccgcc tacggcggct gcggcgccct acgggcttgc tctccgggct
     5041 tcgccctgcg cggtcgctgc gctcccttgc cagcccgtgg atatgtggac gatggccgcg
     5101 agcggccacc ggctggctcg cttcgctcgg cccgtggaca accctgctgg acaagctgat
     5161 ggacaggctg cgcctgccca cgagcttgac cacagggatt gcccaccggc tacccagcct
     5221 tcgaccacat acccaccggc tccaactgcg cggcctgcgg ccttgcccca tcaatttttt
     5281 taattttctc tggggaaaag cctccggcct gcggcctgcg cgcttcgctt gccggttgga
     5341 caccaagtgg aaggcgggtc aaggctcgcg cagcgaccgc gcagcggctt ggccttgacg
     5401 cgcctggaac gacccaagcc tatgcgagtg ggggcagtcg aagggcgaag cccgcccgcc
     5461 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     5521 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     5581 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     5641 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     5701 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     5761 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     5821 tgattgcgca tggtgccgca accgtgcggc acccctaccg catggagata agcatggcca
     5881 cgcagtccag agaaatcggc attcaagcca agaacaagcc cggtcactgg gtgcaaacgg
     5941 aacgcaaagc gcatgaggcg tgggccgggc ttattgcgag gaaacccacg gcggcaatgc
     6001 tgctgcatca cctcgtggcg cagatgggcc accagaacgc cgtggtggtc agccagaaga
     6061 cactttccaa gctcatcgga cgttctttgc ggacggtcca atacgcagtc aaggacttgg
     6121 tggccgagcg ctggatctcc gtcgtgaagc tcaacggccc cggcaccgtg tcggcctacg
     6181 tggtcaatga ccgcgtggcg tggggccagc cccgcgacca gttgcgcctg tcggtgttca
     6241 gtgccgccgt ggtggttgat cacgacgacc aggacgaatc gctgttgggg catggcgacc
     6301 tgcgccgcat cccgaccctg tatccgggcg agcagcaact accgaccggc cccggcgagg
     6361 agccgcccag ccagcccggc attccgggca tggaaccaga cctgccagcc ttgaccgaaa
     6421 cggaggaatg ggaacggcgc gggcagcagc gcctgccgat gcccgatgag ccgtgttttc
     6481 tggacgatgg cgagccgttg gagccgccga cacgggtcac gctgccgcgc cggtagcact
     6541 tgggttgcgc agcaacccgt aagtgcgctg ttccagacta tcggctgtag ccgcctcgcc
     6601 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     6661 acctgaatgg aagccggcgg cacctcgcta acggattcac cgtttttatc aggctctggg
     6721 aggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact
     6781 ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
     6841 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga
     6901 atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc
     6961 aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
     7021 gcagtacggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     7081 acaaaatatt aacgc
//
