LOCUS       sequence-456852-        6425 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of a de novo designed, FRET-based 
            neoLux1.2-mKate2 fusion bioluminescent probe (max. Em=638).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6425)
  AUTHORS   Chen JY, Shi Q, Peng X, De Dieu Habimana J, Wang J, Sobolewski W, 
            Yeh AH
  TITLE     De novo luciferases enable multiplexed bioluminescence imaging
  JOURNAL   Chem. 2024 Nov 12. doi: 10.1016/j.chempr.2024.10.013
REFERENCE   2  (bases 1 to 6425)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Chem. 2024 
            Nov 12. doi: 10.1016/j.chempr.2024.10.013"
COMMENT     Sequence Label: pcDNA3-luxKate
FEATURES             Location/Qualifiers
     source          1..6425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(254..273)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(255..273)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(318..521)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(347..367)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        522..901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1073..1092
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1167..1271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1272..1340,1341..2132)
                     /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     complement(1490..1509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2303..2891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2792..2811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3045..3062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3179..3200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3215..3220,3221..3238,3239..3245)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3253..3269
                     /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     3258..3280
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3277..3293
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3277..3293
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3342..3463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3353..3372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3407..3426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3637..4431)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3748..3767)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4358..4377
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4498..4827)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4512..4647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4566..4585)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4812..4832
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4841..5269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4951..4972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5163..5182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5315..5539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5528..5545
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 ctcgatcttt ctcttggcgc cggtcacggc gctctgcagc ttcttgaacc atgtctcgaa
       61 gccttccttg ctggtgaaca cggtgccgtc ccacaggtga atctccttgc atccggctcc
      121 gaacagcagg tcagcagctg tctcagcgtc tccagcgtca agggcctcgt agaaggcctt
      181 cacgaactcg gcgatctcct cgtcggtcat ggtggcggca agcttaagtt taaacgctag
      241 ccagcttggg tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt
      301 tctctagtta gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc
      361 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
      421 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
      481 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
      541 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
      601 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
      661 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
      721 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
      781 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
      841 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
      901 caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga
      961 ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac
     1021 tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca
     1081 gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt
     1141 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1381 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1441 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1501 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1561 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1621 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1681 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1741 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1801 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1861 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1921 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1981 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2041 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2101 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3001 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3061 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3121 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3181 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3241 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     3301 acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt
     3361 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3421 tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact
     3481 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     3541 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     3601 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3661 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3721 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3781 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3841 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     3901 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3961 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     4021 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     4081 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     4141 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     4201 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     4261 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4321 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4381 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     4441 tcctcatcct gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt
     4501 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga
     4561 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     4621 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     4681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     4741 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4801 ggactttcca caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt
     4861 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4921 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4981 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5041 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5101 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5161 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5221 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc
     5281 ctagagcccc agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca
     5341 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     5401 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     5461 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     5521 tggcaactag aaggcacagt cgaggctgat cagcgggttt aaacgggccc tctagactcg
     5581 agttatctgt gtcccagctt gctaggcagg tcgcagtatc tagcgacggc cacctcgtgc
     5641 tgctcgacgt atgtctcctt gtcggcctct ttaattctct ccagtcttct gtccacgtag
     5701 tacacgccgg gcatcttgag gttcttggcg ggcttcttgc ttctgtatgt tgtcttcagg
     5761 ttgcagataa ggtggccgcc gcccaccagt ttcagggcca tgtcggctct tccttccaat
     5821 cctccgtcag cagggtacag tgtctcggtg ctagcttccc agcccagggt cttcttctgc
     5881 atcacggggc cgttgctggg gaagttcacg cctctgatct tcacgttgta gatcaggcat
     5941 ccgtcttgca ggcttgtgtc ttgggtggcg gtcagcactc caccgtcctc gtatgtggtc
     6001 actctctccc aggtgaagcc ctcggggaag ctctgcttga agaagtcggg gatgccctgg
     6061 gtgtggttga tgaaggtctt gctgccgtac atgaaggatg tagccaggat gtcgaaagcg
     6121 aagggaaggg ggccgccttc cacggccttg attctcatgg tctgggttcc ctcgtaaggc
     6181 tttccctccc cctcgctggt gcacttgaag tggtggttat tcacggtgcc ctccatgtac
     6241 agcttcatgt gcatgttttc cttgatgtaa ggggtggcgg tcaccttcac ctcgaccagc
     6301 tcgttcctct ctctgtcgaa ggtgaaggtg tgctccagct ccacgacgaa cttcttgccg
     6361 gtggccttgt atgtagcagt cagggtcagc ttcaccacgg cctcgccgtc cttgatctcc
     6421 acgct
//
