LOCUS       sequence-248133-        5233 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Prab-3::ssGFP1-10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5233)
  AUTHORS   He S, Cuentas-Condori A, Miller DM
  TITLE     NATF (Native And Tissue-Specific Fluorescence): A Strategy for 
            Bright, Tissue-Specific GFP Labeling of Native Proteins in 
            Caenorhabditis elegans.
  JOURNAL   Genetics. 2019 Apr 5. pii: genetics.119.302063. doi: 
            10.1534/genetics.119.302063.
  PUBMED    30952669
REFERENCE   2  (bases 1 to 5233)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics. 
            2019 Apr 5. pii: genetics.119.302063. doi: 
            10.1534/genetics.119.302063."
COMMENT     Sequence Label: pSH69
FEATURES             Location/Qualifiers
     source          1..5233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..93
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     90..106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     90..106
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    126..146
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1357..1381
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1498..2139
                     /codon_start=1
                     /product="fragment of superfolder GFP consisting of the 
                     first 10 beta-strands"
                     /label=GFP(1-10)
                     /note="associates with GFP11, the 11th beta-strand, to 
                     reconstitute fluorescent GFP (Cabantous et al., 2005; 
                     Kamiyama et al., 2016)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     protein_bind    complement(2141..2165)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2935..2955)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2963..2980)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2963..2979)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2972..2994)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3307..3329
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3367..3385)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3453..3557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3558..3626,3627..4418)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3776..3795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4589..5177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5078..5097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      121 gctatcaact ttgtatagaa aagttgatct tcagatggga gcagtggact gtctatttga
      181 tattcttaaa atgtactcta taaaagatga aactgtaatg accaattccc tacataaata
      241 tttttaacta attggtcgga tatttgggga tcagagaagt acttgttttt gcgatattaa
      301 taatgatata tttttctagt aaaaaaatgt taagcgattt gtgaataatt aagttggctt
      361 gaaaaagatt ttttgatagc tatgtatttt gttaatttaa atcagatttt aaaactttat
      421 tattgactat ttatatatat tatatattat atattttcat ggtttaatca aaatctatta
      481 aattatttct aaaatctcta aatatcgaat tctctaaaaa gcttcatcaa caaaagcatt
      541 tcaattatct aacacaatga gaataattca agaactgaaa tagaaagaaa tctagaaaaa
      601 aaagagagtg aggtgcccta ttaaatcggt gctctcctct ctaacagaag cgcctacacg
      661 cacccaagag agctcttttt ctttcttttt tcggagcaca catacacgca tacactcctc
      721 agaagtgtat agagaaaatg ggagaagaag cgaaagaaga cgaagaagcg gttgtgatgt
      781 atgcatcggg cgtttctatc tctctccgtg agcaacgagc tagtcaaccc atcgatccat
      841 ttttgtgagc acacacagag agagactcaa acacactcac agcttcacac tgagctccaa
      901 gagctctgta gagagaagcc agagtatgct ttcctcgatt ttcctaacgt cttcttcttc
      961 tgcttttttc tttttcccaa cgagttgtag tccactaact tcaatttttc ctccagagaa
     1021 agagttttag agagacagct gataaataca ctagagaaat aaaaagaatg ataaaaattt
     1081 gctattttaa tcagttccct ctcgtttctc taagttttgt tgttgggttg ggaaattaaa
     1141 attggaaaat ggaaataacg tttttttctg tctaatcttg aattctccag acatttctta
     1201 tttccttttt tgttgcaagt ttttcttcct tctctaattt ttaatcaaag atatctcgct
     1261 atgaactttg aatttttgat attttgattc tataaataca taaagtaatc taaatgctct
     1321 tttaaaataa atctacagta gccctatttt cagatgacaa gtttgtacaa aaaagcaggc
     1381 ttaatgttct ccgagttacg aatacttcgc gacatactac ttttatgctt tctatcagtt
     1441 ggaataaatg cgaaatcttc gcatatagaa gatttagact tcacggatca cacaaatatg
     1501 tccaaaggag aagaactgtt taccggtgtt gtgccaattt tggttgaact cgatggtgat
     1561 gtcaacggac ataagttctc agtgagaggc gaaggagaag gtgacgccac cattggaaaa
     1621 ttgactctta aattcatctg tactactggt aaacttcctg taccatggcc gactctcgta
     1681 acaacgctta cgtacggagt tcagtgcttt tcgagatacc cagaccatat gaaaagacat
     1741 gactttttta agtcggctat gcctgaaggt tacgtgcaag aaagaacaat ttcgttcaaa
     1801 gatgatggaa aatataaaac tagagcagtt gttaaatttg aaggagatac tttggttaac
     1861 cgcattgaac tgaaaggaac agattttaaa gaagatggta atattcttgg acacaaactc
     1921 gaatacaatt ttaatagtca taacgtatac atcactgctg ataagcaaaa gaacggaatt
     1981 aaagcgaatt tcacagtacg ccataatgta gaagatggca gtgttcaact tgccgaccat
     2041 taccaacaaa acacccctat tggagacggt ccggtacttc ttcctgataa tcactacctc
     2101 tcaacacaaa cagtcctgag caaagatcca aatgaaaaat acccagcttt cttgtacaaa
     2161 gtgggtgata tctgagctcc gcatcggccg ctgtcatcag atcgccatct cgcgcccgtg
     2221 cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct
     2281 cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt
     2341 cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat
     2401 aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct
     2461 acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat
     2521 catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca
     2581 atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa
     2641 agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct
     2701 gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct
     2761 tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt
     2821 tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg
     2881 gtttttgcct taaatgacag aatacattcc caatatacca aacataactg tttccaactt
     2941 tattatacat agttgataat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3001 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3061 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     3121 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     3181 ctctcagtgc aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3241 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3301 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3361 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3421 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3481 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3541 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3601 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3661 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3721 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3781 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3841 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3901 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3961 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4021 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4081 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4141 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4201 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4261 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gtcagatggt aagccctccc
     4321 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4381 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4441 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4501 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4561 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4621 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4681 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4741 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4801 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4861 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4921 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4981 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5041 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5101 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5161 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gtcttttgct
     5221 ggccttttgc tca
//
