LOCUS       sequence-247876-        5894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An engineered luciferase to enhance the bioluminescence of synthetic
            analog-8pyDTZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  AUTHORS   Yeh HW, Xiong Y, Wu T, Chen M, Ji A, Li X, Ai HW
  TITLE     ATP-Independent Bioluminescent Reporter Variants To Improve in Vivo 
            Imaging.
  JOURNAL   ACS Chem Biol. 2019 Apr 17. doi: 10.1021/acschembio.9b00150.
  PUBMED    30969754
REFERENCE   2  (bases 1 to 5894)
  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 Chem 
            Biol. 2019 Apr 17. doi: 10.1021/acschembio.9b00150."
COMMENT     Sequence Label: pcDNA3-LumiLuc
FEATURES             Location/Qualifiers
     source          1..5894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      664..673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1213..1231)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1214..1231)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1251..1268)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1257..1481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1527..1955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1614..1633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1824..1845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1964..1984)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1969..2298
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2149..2284
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2211..2230
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2365..3159
                     /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(2419..2438)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3029..3048
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3333..3454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3370..3389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3424..3443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3503..3519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3503..3519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3516..3538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3527..3543
                     /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(3551..3557,3558..3575,3576..3581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3596..3617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3734..3751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3905..4493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3985..4004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5525..5629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5704..5723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gccgccacca tggtcttcac tctcggggat tttgttgggg actggcgaca gacagccggc
      721 tacaaccagg ctcaagtcct tgaacaggga ggtttgacca gtttgtttca gaacctcggg
      781 gtgtccgtaa ctccaatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgat
      841 atccatgtca tcatcccgta tgaaggtctg agctgcgacc aaatggccca gatcgaaaaa
      901 atttttaagg tggtataccc tgtggatgat catcacttta aggcgatcct gcactatggc
      961 acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggaca gccgtatgaa
     1021 ggcatcgcca agttcgacgg caaaaagatc acagtaacag ggaccctgtg gaacggcaac
     1081 acaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatt
     1141 aacggagtga ccggctggcg tctgcatgaa cgcattctgg cgtaactcga gcatgcatct
     1201 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     1261 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1321 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1381 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1441 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1501 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1561 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1621 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1681 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1741 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1801 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1861 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1921 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1981 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2041 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2101 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2161 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2221 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2281 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2341 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2401 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2461 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2521 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2581 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2641 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2701 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2761 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2821 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2881 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2941 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3001 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3061 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3121 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3181 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3241 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3301 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3361 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3421 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3481 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3541 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3601 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3661 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3721 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3781 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3841 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4321 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5641 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     5701 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     5761 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     5821 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     5881 cagatatacg cgtt
//
