LOCUS       sequence-134441-        5052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal GFP-2xTEV-Avi vector, for fosmid recombineering.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5052)
  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 5052)
  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: pMB72
FEATURES             Location/Qualifiers
     source          1..5052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(504..532)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    complement(872..905)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        909..956
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(2138..2154)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2138..2154)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(2163..2196)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     2605..2625
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2681..2701
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2705..2725
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2741..2785
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2831..2847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2831..2847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2844..2866)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2855..2871
                     /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(2879..2885,2886..2903,2904..2909))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2924..2945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3062..3079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3233..3821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3313..3332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3992..4783,4784..4852))
                     /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     4615..4634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4853..4957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5025..5043
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagatc cggaggagga ggatctggag gaggaggatc tggaggagga ggaatgagta
      481 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      541 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
      601 cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt ttaaacatat
      661 atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact actttctgtt
      721 atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac tttttcaaga
      781 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
      841 acaagacacg taagtttaaa cagttcggta cgaagttcct atactttcta gagaatagga
      901 acttccctgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
      961 tgaggaacta aacccaggag gcagatcatg agtctgaaag aaaaaacaca atctctgttt
     1021 gccaacgcat ttggctaccc tgccactcac accattcagg cgcctggccg cgtgaatttg
     1081 attggtgaac acaccgacta caacgacggt ttcgttctgc cctgcgcgat tgattatcaa
     1141 accgtgatca gttgtgcacc acgcgatgac cgtaaagttc gcgtgatggc agccgattat
     1201 gaaaatcagc tcgacgagtt ttccctcgat gcgcccattg tcgcacatga aaactatcaa
     1261 tgggctaact acgttcgtgg cgtggtgaaa catctgcaac tgcgtaacaa cagcttcggc
     1321 ggcgtggaca tggtgatcag cggcaatgtg ccgcagggtg ccgggttaag ttcttccgct
     1381 tcactggaag tcgcggtcgg aaccgtattg cagcagcttt atcatctgcc gctggacggc
     1441 gcacaaatcg cgcttaacgg tcaggaagca gaaaaccagt ttgtaggctg taactgcggg
     1501 atcatggatc agctaatttc cgcgctcggc aagaaagatc atgccttgct gatcgattgc
     1561 cgctcactgg ggaccaaagc agtttccatg cccaaaggtg tggctgtcgt catcatcaac
     1621 agtaacttca aacgtaccct ggttggcagc gaatacaaca cccgtcgtga acagtgcgaa
     1681 accggtgcgc gtttcttcca gcagccagcc ctgcgtgatg tcaccattga agagttcaac
     1741 gctgttgcgc atgaactgga cccgatcgtg gcaaaacgcg tgcgtcatat actgactgaa
     1801 aacgcccgca ccgttgaagc tgccagcgcg ctggagcaag gcgacctgaa acgtatgggc
     1861 gagttgatgg cggagtctca tgcctctatg cgcgatgatt tcgaaatcac cgtgccgcaa
     1921 attgacactc tggtagaaat cgtcaaagct gtgattggcg acaaaggtgg cgtacgcatg
     1981 accggcggcg gatttggcgg ctgtatcgtc gcgctgatcc cggaagagct ggtgcctgcc
     2041 gtacagcaag ctgtcgctga acaatatgaa gcaaaaacag gtattaaaga gactttttac
     2101 gtttgtaaac catcacaagg agcaggacag tgctgaggat ccactagttc tagagcggcc
     2161 gcgaagttcc tatactttct agagaatagg aacttctaac taaccataca tatttaaatt
     2221 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
     2281 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
     2341 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
     2401 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     2461 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     2521 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
     2581 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     2641 acatggcatg gatgaactat acaaatctgg aggaggatct gaaaatcttt atttccaagg
     2701 atccgagaac ctttacttcc aaggatctgg aggaggatct ggacttaatg atattttcga
     2761 agctcagaag attgaatggc atgaataaag atctgtcgac ctgcaggcat gcaagcttgg
     2821 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2881 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2941 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3001 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3061 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3121 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3181 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3241 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3301 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3361 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3421 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3481 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3541 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3601 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3661 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3721 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3781 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3841 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3901 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3961 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4021 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4081 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4141 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4201 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4261 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4321 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4381 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4441 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4501 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4561 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4621 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4681 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4741 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4801 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4861 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4921 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4981 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5041 ggccctttcg tc
//
