LOCUS       sequence-252849-        6331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of luxB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6331)
  AUTHORS   Gregor C, Pape JK, Gwosch KC, Gilat T, Sahl SJ, Hell SW
  TITLE     Autonomous bioluminescence imaging of single mammalian cells with 
            the bacterial bioluminescence system.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Dec 2. pii: 1913616116. doi: 
            10.1073/pnas.1913616116.
  PUBMED    31792180
REFERENCE   2  (bases 1 to 6331)
  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: "Proc Natl 
            Acad Sci U S A. 2019 Dec 2. pii: 1913616116. doi: 
            10.1073/pnas.1913616116."
COMMENT     Sequence Label: co luxB pcDNA3.1(+)
FEATURES             Location/Qualifiers
     source          1..6331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1922..1939)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1928..2152
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2198..2626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2285..2304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2495..2516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2635..2655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2640..2969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2820..2955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2882..2901
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3036..3830
                     /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(3090..3109)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3700..3719
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4004..4125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4041..4060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4095..4114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4174..4190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4174..4190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4187..4209)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4198..4214
                     /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(4222..4228,4229..4246,4247..4252))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4267..4288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4405..4422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4576..5164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4656..4675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5335..6126,6127..6195))
                     /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     5958..5977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6196..6300)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atgaagttcg gcctgttctt cctgaacttc atcaacagca ccaccgtgca ggagcagagc
      961 atcgtgcgca tgcaggagat caccgagtac gtggacaagc tgaacttcga gcagatcctg
     1021 gtgtacgaga accacttcag cgacaacggc gtggtgggcg cccccctgac cgtgagcggc
     1081 ttcctgctgg gcctgaccga gaagatcaag atcggcagcc tgaaccacat catcaccacc
     1141 caccaccccg tgcgcatcgc cgaggaggcc tgcctgctgg accagctgag cgagggccgc
     1201 ttcatcctgg gcttcagcga ctgcgagaag aaggacgaga tgcacttctt caaccgcccc
     1261 gtggagtacc agcagcagct gttcgaggag tgctacgaga tcatcaacga cgccctgacc
     1321 accggctact gcaaccccga caacgacttc tacagcttcc ccaagatcag cgtgaacccc
     1381 cacgcctaca cccccggcgg cccccgcaag tacgtgaccg ccaccagcca ccacatcgtg
     1441 gagtgggccg ccaagaaggg catccccctg atcttcaagt gggacgacag caacgacgtg
     1501 cgctacgagt acgccgagcg ctacaaggcc gtggccgaca agtacgacgt ggacctgagc
     1561 gagatcgacc accagctgat gatcctggtg aactacaacg aggacagcaa caaggccaag
     1621 caggagaccc gcgccttcat cagcgactac gtgctggaga tgcaccccaa cgagaacttc
     1681 gagaacaagc tggaggagat catcgccgag aacgccgtgg gcaactacac cgagtgcatc
     1741 accgccgcca agctggccat cgagaagtgc ggcgccaaga gcgtgctgct gagcttcgag
     1801 cccatgaacg acctgatgag ccagaagaac gtgatcaaca tcgtggacga caacatcaag
     1861 aagtaccaca tggagtacac ctgactcgag tctagagggc ccgtttaaac ccgctgatca
     1921 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1981 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2041 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2101 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2161 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2221 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2281 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2341 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2401 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2461 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2521 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2581 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2641 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2701 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2761 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2821 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2881 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     2941 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3001 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3061 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3121 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3181 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3241 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3301 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3361 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3421 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3481 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3541 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3601 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3661 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3721 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3781 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3841 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3901 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3961 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4021 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4081 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4141 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4201 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4261 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4321 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4381 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4441 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4501 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4561 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4621 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4681 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4741 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4801 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4861 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4921 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4981 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5041 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5101 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5161 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5221 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5281 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5341 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5401 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5461 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5521 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5581 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5641 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5701 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5761 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5821 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5881 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5941 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6001 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6061 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6121 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6181 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6241 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6301 cacatttccc cgaaaagtgc cacctgacgt c
//
