LOCUS       sequence-456855-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for de novo luciferase, neoLux1.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  AUTHORS   Chen JY-H, Shi Q, Peng X, Habimana JdD, Wang J, Sobolewski W, Yeh 
            AH-W
  TITLE     De novo luciferases enable multiplexed bioluminescence imaging.
  JOURNAL   Chem. 2025 Mar 13;11(3):102346. doi: 10.1016/j.chempr.2024.10.013. 
            Epub 2024 Nov 12.
  PUBMED    40417169
REFERENCE   2  (bases 1 to 5612)
  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: "Chem."; 
            date: "2025-03-13"; volume: "11(3):102346. doi"; pages: " 
            10.1016/j.chempr.2024.10.013. Epub 2024 Nov 12"
COMMENT     Sequence Label: pET29-neoLux1.2
FEATURES             Location/Qualifiers
     source          1..5612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1524..1715
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1709..1731)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1817..1956
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1872..1891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1971..1988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2142..2730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2222..2241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2841..2860)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2852..3667
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2852..3667
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2925..2944)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3759..4214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3896..3917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4108..4127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4251..4298)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4294..4312
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4368..4385)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4774..4796
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4811..4835
                     /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)."
     primer_bind     complement(4835..4854)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4836..4854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4942..4961)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5167..5244
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5245..5612,1..715)
                     /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"
     primer_bind     complement(5264..5283)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
       61 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
      121 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
      181 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
      241 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
      301 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
      361 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
      421 cgttggtgcg gacatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
      481 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
      541 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
      601 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      661 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      721 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
      781 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
      841 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
      901 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
      961 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     1021 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     1081 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     1141 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     1201 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     1261 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     1321 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     1381 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     1441 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     1501 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     1561 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     1621 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     1681 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     1741 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     1801 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     1861 gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata
     1921 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     1981 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2041 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2101 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2161 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2221 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2281 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2341 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2401 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2461 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2521 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2581 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2641 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2701 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     2821 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     2881 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2941 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     3001 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3061 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3121 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     3181 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3241 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3301 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3361 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3421 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3481 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3541 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3601 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3661 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3721 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     3781 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3841 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3901 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3961 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4021 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4081 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4141 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4201 gctacagggc gcgtcccatt cgccaatccg gatatagttc ctcctttcag caaaaaaccc
     4261 ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca
     4321 gccaactcag cttcctttcg ggctttgtta gcagccggat ctcattagtg gtggtggtgg
     4381 tggtgggaac ctagtggtgt tgctgtgact ttcacttcta ctagttcgtt tctttctctg
     4441 tcgaatgtga atgtgtgttc tagttctact acgaattttt ttcctgttgc tttgtatgtt
     4501 gctgttagtg ttagttttac tactgcttct ccgtctttta tttctacgct ttctattttt
     4561 ctttttgctc ctgttactgc gctttgtagt tttttgaacc atgtttcgaa tccttctttg
     4621 cttgtgaata ctgttccgtc ccataggtgt atttctttgc atcctgctcc gaatagtagg
     4681 tctgctgctg tttctgcgtc tcctgcgtcg agtgcttcgt agaatgcttt tacgaattct
     4741 gctatttctt cgtctgttcc tgacatatgt atatctcctt cttaaagtta aacaaaatta
     4801 tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
     4861 cgggatcgag atcgatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg
     4921 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     4981 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     5041 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     5101 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag
     5161 atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     5221 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     5281 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     5341 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     5401 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     5461 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     5521 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     5581 aatcttctcg cgcaacgcgt cagtgggctg at
//
