LOCUS       sequence-140201-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LucC-hDib1 fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  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 6370)
  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: pSplit-CO-hDib1
FEATURES             Location/Qualifiers
     source          1..6370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /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(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2493..2510
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2994..3017)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3017..3026
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3032..3058
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHYHHHH"
     primer_bind     complement(3552..3571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3553..3571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3616..3819)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3645..3665)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3820..4199
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4371..4390
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4465..4569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4570..4638,4639..5430)
                     /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(4788..4807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6343..6360
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgatca ctcgaggtag cggtacttgg tggagtagtc cttgggggac
     2581 accaccaggc cgcggccttt gcgggccccg cggtacaccg tctcgatgat gtccaccatc
     2641 tcctgcttgt cctccatggc ccagttaatc ttgttgttgt tgccagtccc caagtcaatc
     2701 atgatgtgct tgttcctgaa gaaaaacatg acagtacatg gatcgtataa ctcatacatt
     2761 ttgttgaagt caggcacttc tgtaatatcc acaagataaa taactgcaaa atttttaacc
     2821 ttctcggcga tgctgtacag gacctcgtcc atcttcatgc acgtaggatc ccagtcgtgg
     2881 ccgaagcgga tgacgaccac gcggtcctcc tccgagagga tggcctggtc cacctgccag
     2941 ccgttgtgca ggtgcgggag catgtacgac atgaattcaa ctagagctag ttccttgtca
     3001 tcgtcgtcct tgtagtccat ggtggcggat ccaccaccac cactaccacc accaccaccg
     3061 gtacccaatt tggactttcc gcccttcttg gcctttatga ggatctctct gatttttctt
     3121 gcgtcgagtt ttccggtaag acctttcggt acttcgtcca caaacacaac tcctccgcgc
     3181 aactttttcg cggttgttac ttgactggcg acgtaatcca cgatctcttt ttccgtcatc
     3241 gtctttccgt gctccaaaac aacaacggcg gcgggaagtt caccggcgtc atcgtcggga
     3301 agacctgcca cgcccgcgtc gaagatgttg gggtgttgta acaatatcga ttccaattca
     3361 gcgggggcca cctgatatcc tttgtattta attaaagact tcaagcggtc aactatgaag
     3421 aagtgttcgt cttcgtccca gtaagctatg tctccagaat gtagccatcc atccttgtca
     3481 atcaaggcgt tggtcgcttc cggattgttt acataaccgg acataatcat aggtcccata
     3541 agcttgggtc tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc
     3601 tctagttagc cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     3661 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     3721 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     3781 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     3841 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     3901 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     3961 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     4021 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     4081 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     4141 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
     4201 acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt
     4261 cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta
     4321 ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga
     4381 gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca
     4441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4801 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6361 gaagcggaag
//
