LOCUS       sequence-370974-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LgBiT via sigma 70 promoter in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  AUTHORS   Sato W, Rasmussen M, Deich C, Engelhart AE, Adamala KP
  TITLE     Expanding luciferase reporter systems for cell-free protein 
            expression.
  JOURNAL   Sci Rep. 2022 Jul 7;12(1):11489. doi: 10.1038/s41598-022-15624-6.
  PUBMED    35798760
REFERENCE   2  (bases 1 to 5787)
  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: "Sci Rep."; 
            date: "2022-07-7"; volume: "12(1):11489. doi"; pages: " 
            10.1038/s41598-022-15624-6"
COMMENT     Sequence Label: pLumi-LgBiT
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        887..934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     1601..1621
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     rep_origin      complement(1752..2297)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1832..1851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2461..2547)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2700..2717
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(3262..4053,4054..4122))
                     /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     3885..3904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4123..4227)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4295..4313
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taataactat ctgaagcaac gaattaatga tgagaacgaa atatcaatag tcaccgaagc
       61 aatgaatgaa gcacttcgta aacttgagca acgtgtatta aatacgctca atgaattttc
      121 tggttatact catgttatgg ttataggcgg tggcgcagaa ttaatatgcg atgcagtaaa
      181 aaaacacaca cagattcgtg atgaacgttt tttcaaaacc aataactctc aatatgattt
      241 agttaacggt atgtatctca taggtaatta atgatggaca agcgcagaac cattgccttc
      301 aaactaaatc cagatgtaaa tcaaacagat aaaattgttt gtgatacact ggacagtatc
      361 ccgcaagggg aacgaagccg ccttaaccgg gccgcactga cggcaggtct ggccttatac
      421 agacaagatc cccggacccc tttcctttta tgtgagctgc tgacgaaaga aaccacattt
      481 tcagatatcg tgaatatatt gagatcgcta tttccaaaag agatggccga ttttaattct
      541 tcaatagtca ctcaatcctc ttcacaacaa gagcaaaaaa gtgatgaaga gaccaaaaaa
      601 aatgcgatga agctaataaa ttaattcaat tattattgag ttccctttat ccactatcag
      661 gctggataaa gggaactcaa tcaagttatt ttcttaccag tcattacata atcgttatta
      721 tgaaataatc gtttgcactg tctctgttat tcaggcaatt tcaataaagg cacttgctca
      781 cgctctgtca ttttctgaaa ctctcatgtg gaattcctgc agggcatgcc ccgggcatat
      841 gaagcttggt accgggatcc gcggccgcgt ttaaacccat ggacgtgttg acaattaatc
      901 atcggcatag tatatcggca tagtataata cgactcaggg cccactagtg gtacccgggg
      961 atcctctaga gtcgacctgc aggtcgacgg atccggggaa ttcaggcttg gaggatacgt
     1021 atggtcttca cactcgaaga tttcgttggg gactgggaac agacagccgc gtacaacctg
     1081 gaccaagtcc ttgaacaggg aggtgtgtcc agtttgctgc agaatctcgc ggtgtccgta
     1141 actccgatcc aaaggattgt ccgcagcggt gaaaatgcgc tgaagatcga catccatgtc
     1201 atcatcccgt atgaaggtct gagcgcggac caaatggcac agatcgaaga agtgtttaag
     1261 gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgccgtatgg cacactggta
     1321 atcgacgggg ttacgccgaa catgctgaac tatttcggac ggccgtatga aggcatcgcc
     1381 gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc
     1441 gacgagcgcc tgatcacccc cgacggctcc atgctgttcc gagtaaccat caacagctga
     1501 cctgcaggca tgcaagcttg cggccgcggg ccccatggat cgatagctgg tcgacacaat
     1561 ctgccctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa
     1621 ctgctgctgg gattacacat ggcatggatg agctctacaa ataatgagct agcccgccta
     1681 atgagcgggc ttttttttct cggcctagga gatacttaac agggaagtga gagggccgcg
     1741 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     1801 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     1861 tcccccgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     1921 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     1981 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2041 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2101 atttagagga gttagtcttg aagccatgcg ccggttaagg ctaaactgaa aggacaagtt
     2161 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     2221 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     2281 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc taggatctaa
     2341 aacactaggc cccgtcagga tggccttctg cttaatttga tgcctggcag tttatggcgg
     2401 gcgtcctgcc cgccaccctc cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg
     2461 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     2521 tttcgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcatggggag
     2581 accccacact accatcggcg ctacggcgtt tcacttctga gttcggcatg gggtcaggtg
     2641 ggaccaccgc gctactgccg ccaggcaaat tctgttttat cagaccgctt ctgcgttctg
     2701 atttaatctg tatcaggctg aaaatcttct ctcatccgcc aaaacagcca agctggatct
     2761 ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt
     2821 tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta
     2881 caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac
     2941 gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt
     3001 tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt
     3061 cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct
     3121 tactgtcaac cggatctaaa acactagtta agggattttg gtcatgagat tatcaaaaag
     3181 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3241 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3301 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3361 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3421 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3481 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3541 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3601 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatggtc
     3661 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3721 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3781 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3841 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3901 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3961 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4021 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4081 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4141 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4201 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4261 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4321 tcttctagac aacatcagca aggagaaagg ggctaccggc gaaccagcag cccctttata
     4381 aaggcgcttc agtagtcaga ccagcatcag tcctgaaaag gcgggcctgc gcccgcctcc
     4441 aggttgctac ttaccggatt cgtaagccat gaaagccgcc acctccctgt gtccgtctct
     4501 gtaacgaatc tcgcacagcg attttcgtgt cagataagtg aatatcaaca gtgtgagaca
     4561 cacgatcaac acacaccaga caagggaact tcgtggtagt ttcatggcct tcttctcctt
     4621 gcgcaaagcg cggtaagagg ctatcctgat gtggactaga catagggatg cctcgtggtg
     4681 gttaatgaaa attaacttac tacggggcta tcttctttct gccacacaac acggcaacaa
     4741 accaccttca cgtcatgagg cagaaagcct caagcgccgg gcacatcata gcccatatac
     4801 ctgcacgctg accacactca ctttccctga aaataatccg ctcattcaga ccgttcacgg
     4861 gaaatccgtg tgattgttgc cgcatcacgc tgcctcccgg agtttgtctc gagcactttt
     4921 gttacccgcc aaacaaaacc caaaaacaac tcatacccaa cccaataaaa caccaaaaca
     4981 agacaaataa tcattgattg atggttgaaa tggggtaaac ttgacaaaca aacccactta
     5041 aaacccaaaa catacccaaa cacacaccaa aaaaacacca taaggagttt tataaatgtt
     5101 ggtattcatt gatgacggtt caacaaacat caaactacag tagcaggaaa gcgacggaac
     5161 aattaaacag cacattagcc cgaacagctt caaacgcgag tgggcagtct cttttggtga
     5221 taaaaaggtc tttaactaca cactgaacgg cgaacagtat tcatttgatc caatcagccc
     5281 ggatgctgta gtcacaacca atatcgcatg gcaatacagc gacgttaatg tcgttgcagt
     5341 gcatcacgcc ttactgacca gtggtctgcc ggtaagcgaa gtggatattg tttgcacact
     5401 tcctctgaca gagtattacg acagaaataa ccaacccaat acggaaaata ttgagcgtaa
     5461 gaaagcaaac ttccggaaaa aaattacatt aaatggcggg gatacattca caataaaaga
     5521 tgtaaaagtc atgcctgaat ctataccggc aggttatgaa gttctacaag aactggatga
     5581 gttagattct ttattaatta tagatctcgg gggcaccaca ttagatattt ctcaggtaat
     5641 ggggaaatta tcggggatca gtaaaatata cggagactca tctcttggtg tctctctggt
     5701 tacatctgca gtaaaagatg ccctttctct tgcgagaaca aaaggaagta gctatcttgc
     5761 tgacgatata atcattcaca gaaaaga
//
