LOCUS       sequence-424721-        6620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LucN alone in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  AUTHORS   Maita H, Tomita K, Ariga H
  TITLE     A split luciferase-based reporter for detection of a cellular 
            macromolecular complex.
  JOURNAL   Anal Biochem. 2014 Feb 3. pii: S0003-2697(14)00035-9. doi: 
            10.1016/j.ab.2014.01.015.
  PUBMED    24503441
REFERENCE   2  (bases 1 to 6620)
  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: "Anal 
            Biochem. 2014 Feb 3. pii: S0003-2697(14)00035-9. doi: 
            10.1016/j.ab.2014.01.015."
COMMENT     Sequence Label: pcDNA3-FLAG-Luc-N
FEATURES             Location/Qualifiers
     source          1..6620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..79)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        64..393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     306..325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             460..1254
                     /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(514..533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1124..1143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1428..1549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1465..1484)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1519..1538
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1598..1614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1598..1614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1611..1633)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1622..1638
                     /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(1646..1652,1653..1670,1671..1676))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1691..1712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1829..1846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2000..2588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2080..2099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2759..3550,3551..3619))
                     /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     3382..3401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3620..3724)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3799..3818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3990..4369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4370..4573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4524..4544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4618..4637
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4618..4636
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4653..4676
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4770..4788)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(5928..5946)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5929..5946)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5966..5983)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5972..6196
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6242..6620,1..50)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6329..6348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6539..6560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
       61 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
      361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
      421 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
      481 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
      541 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
      601 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
      661 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
      721 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
      781 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
      841 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
      901 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
      961 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     1021 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     1081 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     1141 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     1201 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     1261 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     1321 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     1381 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     1441 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1501 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     1561 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1621 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1681 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1741 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1801 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1861 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1921 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1981 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2041 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2101 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2161 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2221 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2281 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2341 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2401 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2461 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2521 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2581 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2641 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2701 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2761 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2821 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2881 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2941 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3001 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3061 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3361 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3601 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3721 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     3781 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     3841 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     3901 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     3961 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     4021 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     4081 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4141 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4201 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4261 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     4321 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     4381 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     4441 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     4501 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     4561 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     4621 tacgactcac tatagggaga cccaagctta tggactacaa ggacgacgat gacaaggaac
     4681 tagctctagt tgaattcgga tccatggaag acgccaaaaa cataaagaaa ggcccggcgc
     4741 cattctatcc tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat
     4801 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt
     4861 acgcggaata cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga
     4921 atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt
     4981 tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg
     5041 aattgctcaa cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt
     5101 tgcaaaaaat tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg
     5161 attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac
     5221 ctcccggttt taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg
     5281 cactgataat gaactcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata
     5341 gaactgcctg cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc
     5401 cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac
     5461 tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt
     5521 ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt
     5581 cattcttcgc caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg
     5641 cttctggggg cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc
     5701 ttccagggat acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac
     5761 ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg
     5821 ttgtggatct ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca
     5881 gaggacctat gattatgtga ctcgagcatg catctagagg gccctattct atagtgtcac
     5941 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6001 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6061 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6121 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     6181 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     6241 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6601 taagggattt tgccgatttc
//
