LOCUS       sequence-191039-        3789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For autolysate production in lacZ deficient (lac operon deletion) E.
            coli BL21-Gold-dLac (DE3) strain: constitutively expresses phage 
            lambda endolysin (gene R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3789)
  AUTHORS   Didovyk A, Tonooka T, Tsimring L, Hasty J
  TITLE     Rapid and scalable preparation of bacterial lysates for cell-free 
            gene expression.
  JOURNAL   ACS Synth Biol. 2017 Aug 10. doi: 10.1021/acssynbio.7b00253.
  PUBMED    28795570
REFERENCE   2  (bases 1 to 3789)
  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: "ACS Synth 
            Biol. 2017 Aug 10. doi: 10.1021/acssynbio.7b00253."
COMMENT     Sequence Label: pAD-LyseR (in lacZ deficient strain)
FEATURES             Location/Qualifiers
     source          1..3789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..136
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(647..664)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      817..903
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      995..1022
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1041..1132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1133..1201,1202..1993)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1351..1370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2035..2490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2122..2141)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2332..2353
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2601..3189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3090..3109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3343..3360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3526..3548
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3686..3705
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgataggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
       61 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      121 ttgaaaaagg aagagtatgg tagaaatcaa taatcaacgt aaggcgttcc tcgatatgct
      181 ggcgtggtcg gagggaactg ataacggacg tcagaaaacc agaaatcatg gttatgacgt
      241 cattgtaggc ggagagctat ttactgatta ctccgatcac cctcgcaaac ttgtcacgct
      301 aaacccaaaa ctcaaatcaa caggcgccgg acgctaccag cttctttccc gttggtggga
      361 tgcctaccgc aagcagcttg gcctgaaaga cttctctccg aaaagtcagg acgctgtggc
      421 attgcagcag attaaggagc gtggcgcttt acctatgatt gatcgtggtg atatccgtca
      481 ggcaatcgac cgttgcagca atatctgggc ttcactgccg ggcgctggtt atggtcagtt
      541 cgagcataag gctgacagcc tgattgcaaa attcaaagaa gcgggcggaa cggtcagaga
      601 gattgatgta tgataactgt tttggcggat gagagaagat tttcagcctg atacagatta
      661 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
      721 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
      781 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
      841 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
      901 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
      961 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     1021 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct
     1081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     1381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     2041 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2101 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2161 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2221 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2281 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2341 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     2401 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2461 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     2521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3361 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3421 cgcatagggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3481 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3541 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca
     3601 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     3661 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     3721 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggagctgc tcatgtttga
     3781 cagcttatc
//
