LOCUS       sequence-149917-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes E.coli biotin-ligase under neuronal promotor for C.elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  AUTHORS   Waaijers S, Munoz J, Berends C, Ramalho JJ, Goerdayal SS, Low TY, 
            Zoumaro-Djayoon AD, Hoffmann M, Koorman T, Tas RP, Harterink M, 
            Seelk S, Kerver J, Hoogenraad CC, Bossinger O, Tursun B, van den 
            Heuvel S, Heck AJ, Boxem M
  TITLE     A tissue-specific protein purification approach in Caenorhabditis 
            elegans identifies novel interaction partners of DLG-1/Discs large.
  JOURNAL   BMC Biol. 2016 Aug 9;14:66. doi: 10.1186/s12915-016-0286-x.
  PUBMED    27506200
REFERENCE   2  (bases 1 to 8349)
  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: "BMC Biol.";
            date: "2016-08-9"; volume: "14:66. doi"; pages: " 
            10.1186/s12915-016-0286-x"
COMMENT     Sequence Label: rgef-1p::NmBirA
FEATURES             Location/Qualifiers
     source          1..8349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3586..3606
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3631..3660
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3667..3696
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5789..5811)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5911..5930
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6008..6026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6094..6198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6199..6267,6268..7059)
                     /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(6417..6436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7230..7818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7719..7738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7972..7989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8106..8127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8142..8147,8148..8165,8166..8172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8180..8196
                     /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     8185..8207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8204..8220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8204..8220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcagcg tttccgatac ccccttatat cagcacacat tcagtcatca
       61 taccacaaaa tgttgtgata gtgataggca aaactaccgt atttcctcta ttaatcttgc
      121 atgcaagact aattttcgat taacccgtag gggtgcaaga ctaatagaga ctgcaagact
      181 attagaggct gaaatactaa ttttcgtatg ctcaataatt ttggaaattg gcctattttt
      241 tgtagaaact tgataccgtt taaacaagga aaaatacaca ctttttaata tttcatcaat
      301 aatttgaacg attttgtgat tttaagttca atttgcccaa aaaaagacaa tttttctgac
      361 gttaccgcaa agtaatgctg gtcaggcaaa aagagcggtg caaaaatata agagactgca
      421 atactaatag aggaaatacg gtaattgaat tttagtgaaa cttgcgacag ttttcctcat
      481 ttttttgtta ttccggcata cgagtgtggc atacgagtga ggtcatcttt tgtttcttcc
      541 gtttcttcat cgcttttgaa aaaaaatgtt gaagaaaact tcgtagccga tagccggata
      601 atgtttggca cggcgttcca acatatatca ttggaaattt tttaagattt ttgcgaaaaa
      661 tcacattctt cacgatgaga acacgttatt gaaggaatat aaatcaagaa taacatatag
      721 ttatatttct ctattacttt aacgttaaat atgagcaaat ttgagcattt tgattgcgat
      781 gaaaagcaga atcgagtcaa ctgaaatccg ttcaaaaacc taagctcggt tcgcccgaaa
      841 gtcgattttt ccaatagccg aacagcactg cttctttatt ttccaggtat ttttggggtg
      901 taagaattat atcttctttg tttgggtatt tacgtttccg aattcccttt cttgatctgt
      961 cgctattttt cagaaaccag aacagtttat cctttttcat gagaacctga atagctcaaa
     1021 aacccgttta atttcttttc atcatctcat ttgaaacttt cctagtcttc ttaatgattt
     1081 ccaggctcct cctacatttg tatctcaaga tgcatacact ttatttccgc tccaattcga
     1141 atcatctact tctctttttt cttcaacttt ctactttttt ccattctttt ctttcaaatg
     1201 ttcgacgtct tcaagtgact gctcttcatt ctcctcattt ttctcgaatt tcattcttgt
     1261 ctcttttttg ctcaaaaatg gaaaatgaaa gtgacgtgag atttggacgg cgggacacgg
     1321 gggcagtaga agcagcaaaa aggagagaaa gaggacacaa ataagaagaa cgaattcaaa
     1381 aataagcgga gaggagctat ttccgtcaat tctacctccc caatcttcat caattcggct
     1441 caattgaatg acgtcacagg gagataaacc gtttggatga gcgccgtcca tcactgacgc
     1501 catcccgttt gggacaagaa aagagaaaaa agagcacaaa gtttttggtg acggatcttg
     1561 tcaatcatat gaaagttgtt ctgattgatt gtcagttttt ttcctacttt tttggattct
     1621 atccacttct gaacttttga caagtttcaa actttctgaa tcattttcta tgcattttcc
     1681 tggaattctt tttatgtaaa atatgaaata gaatgttttt gaattcaagt tctgcttttt
     1741 tcttcttttt tgttctgttc gggcttgggt atgctttttt taaaaattat tttgcacatc
     1801 gaccaataag tgcgcaactt ataaaattaa tttatttttg ttaatttttg aaatacttgt
     1861 attgctttaa gtgatctgac ctcgccctga gctttccacg tagttatcaa atacaaatcc
     1921 tccaagggta acgtacctat attactgatc tttataataa ctttatcacc tgtccagttc
     1981 cagaggaatt ctgttaagct tataggcaca gaaggagtca tttctgctgg tttttaatga
     2041 tccaaatctt tatttcaagt aaaaaactga acacttgcga ataaaactat cagattaacc
     2101 attcaccaaa aatgtgtttg aatctaaaac tttctcagta ttccaaatat agaaataaat
     2161 aaccacgaca ttgctaaaat ctgtctgaat tgtgtactcc ttaccgtgaa gtaataatgg
     2221 atataatgaa tcgtttgaaa tgaatgatca gcacattttt ggtgaaagat cacaaataag
     2281 gaataagcga cggaaaataa aacgattatt tcggatcaaa aatttgttga agatcatata
     2341 cactcgagac caagattatt ctagacaatt ttcaaattgg cttctttgtt tgcaaatctt
     2401 cataataatc tgtgaagttt ggaaatttga atttttaatc ttttttcata gattatagtt
     2461 tttatttctt tgcaaaacta tattaaaaac cgatgcattg ttttagggaa attaatgagc
     2521 cttttgttca acactaaaaa caataaaatt aaaattttgg cttcatcatt tgaccttttt
     2581 taagttcgaa aacttttctc gtatttctga accgccaatt ttttcacaca tctctagact
     2641 tttggtgccc gttccagaaa gttaagtaat tgctattcta agaaagttct caacattgtt
     2701 ttttagttct gattgaattc tgatgttcca ggaatatatt ttaaatttaa tattctttgc
     2761 actatttcta tactaactaa ataataaata gtctaagtat gttcaatgag aacaaaaaag
     2821 ctctatctat atttttctat cctatttcat ttatatcctt ttcattttga actcaccagc
     2881 ttctcttctt cttcttgtcc atatacttct taaaagtctc gaacttctct ctctcactct
     2941 ccatcaattt ctttatggct accgcttgcg ctctgccgct tccgaagaaa gaaggcgttg
     3001 gcagagctct caattctatt ttttctcgcc gtaggctatc gatttctcag cttctcttct
     3061 ctcttcctca ctccaccacc accacttcgg gcttcttctt ttcttcattt tcgtcttctt
     3121 cttcatcctc ttcttttttt ttcagttccc cctcactcct ccccttatat gcgcgtgcga
     3181 gaggggtgca aaagcagcgc gcatccgaga attgaaacga gaaaacggag acgcagcagc
     3241 agttcgtcct cagaaaaata gccagtaaaa agagaaaaag atagagagag acctatcgca
     3301 ttttattttc aaattgcaat tcctgcaatt catgtgtgcg tgtgcccatt ttcaattctt
     3361 cccgttattt ttcagaccat taccaacgtt ctttctagat cttgagacaa tcttccttct
     3421 gctcaatcgt tcgtcgtgaa gacggcatcg acgacgacga cgatcagcag taaaggatcc
     3481 ccgggattgg ccaaaggacc caaaggtatg tttcgaatga tactaacata acatagaaca
     3541 ttttcaggag gacccttgag ggtaccggta gaaaaaatga ccgctccaaa gaagaaacgc
     3601 aaagtaccga gctcagaaaa gaccggtatg gagcagaagc tcatctctga ggaagacctc
     3661 ggaggagagc agaaactcat ctctgaagag gacctcatga aggataatac tgttccattg
     3721 aagttgattg ctttgcttgc aaatggagaa tttcattctg gagaacaact tggagagacg
     3781 ttgggaatgt cgcgtgctgc aattaacaag catattcaaa cagtaagttt aaacagttcg
     3841 gtactaacta accatacata tttaaatttt cagttgagag actggggagt tgacgttttt
     3901 acggttccag gaaagggata ttcgcttcca gagccaattc agctcttgaa tgcaaagcaa
     3961 attttgggac agcttgatgg tggtagtgtt gcagtacttc cggttataga tagtacaaat
     4021 cagtacttgc ttgatcgaat aggagaattg aaatcaggag atgcttgcat cgctgagtat
     4081 cagcaggctg gacgaggacg aagaggacgt aagtggttct gtaagtttaa acatatatat
     4141 actaactaac cctgattatt taaattttca gcgcctttcg gagccaatct ctatttgagt
     4201 atgttctggc gtcttgaaca aggtccggcc gctgctatcg gactttcatt ggttattgga
     4261 attgtgatgg cagaagttct ccgaaaactt ggtgcagata aggtgagagt caagtggcct
     4321 aacgatcttt atcttcaaga tagaaaattg gccggaatat tggtaagttt aaacatatat
     4381 atactaacta accctgatta tttaaatttt caggttgagc ttactggaaa gacgggagat
     4441 gctgctcaaa ttgtgattgg agctggtatc aatatggcaa tgagaagagt agaagagtct
     4501 gttgttaatc aaggttggat cacacttcaa gaggcaggaa tcaaccttga tagaaatacg
     4561 ttggccgcca tgcttatccg agaattgcga gctgcactcg aactcttcga acaagaaggt
     4621 cttgtaagtt taaacatata tatactaact aaccctgatt atttaaattt tcaggcccct
     4681 tacctcagta gatgggagaa acttgacaat tttatcaatc gaccagtgaa gttgataatt
     4741 ggagataagg aaatttttgg tatcagtcga ggaattgata agcaaggagc tcttctcctt
     4801 gagcaagatg gaatcattaa accatggatg ggtggagaga tttctttgcg atctgctgaa
     4861 aagtaagaat tccaactgag cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa
     4921 taataggggc cgctgtcatc agagtaagtt taaactgagt tctactaact aacgagtaat
     4981 atttaaattt tcagcatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa
     5041 catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa
     5101 acttcttctt aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg
     5161 tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc
     5221 ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt
     5281 tttttttact tctgataaat tttttttgaa acatcataga aaaaaccgca cacaaaatac
     5341 cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc
     5401 ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt
     5461 caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg ggtttcccct
     5521 attgtttgtc aagagtttcg aggacggcgt ttttcttgct aaaatcacaa gtattgatga
     5581 gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga aggtgagtag
     5641 aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa tgacagaata
     5701 cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg ccctttcgtc
     5761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5881 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5941 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcggc
     6001 cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6061 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6181 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6241 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6301 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6361 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6421 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6481 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6541 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6601 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6661 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6721 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6781 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6841 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6901 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6961 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7021 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     7381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7621 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7921 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7981 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     8041 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     8101 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     8161 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     8221 catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat tctcatttaa
     8281 attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct aacaacttgg
     8341 aaatgaaat
//
