LOCUS       sequence-111543-        7135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses gfp::h2b in C. elegans cells from the ubiquitous eef-1A.1 
            (eft-3) promoter and contains a C. briggsae unc-119 rescuing 
            fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7135)
  AUTHORS   Katic I, Xu L, Ciosk R
  TITLE     CRISPR/Cas9 Genome Editing in Caenorhabditis elegans: Evaluation of 
            Templates for Homology-Mediated Repair and Knock-Ins by 
            Homology-Independent DNA Repair.
  JOURNAL   G3 (Bethesda). 2015 Jun 3. pii: g3.115.019273. doi: 
            10.1534/g3.115.019273.
  PUBMED    26044730
REFERENCE   2  (bases 1 to 7135)
  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: "G3 
            (Bethesda). 2015 Jun 3. pii: g3.115.019273. doi: 
            10.1534/g3.115.019273."
COMMENT     Sequence Label: pIK137
FEATURES             Location/Qualifiers
     source          1..7135
                     /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"
     promoter        450..1049
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     protein_bind    1058..1082
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1115..1143)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1891..1911
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2372..2396)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2401..2724
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     CDS             4705..4716
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4914..4930)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4914..4930)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4927..4949)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4938..4954
                     /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(4962..4968,4969..4986,4987..4992))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5007..5028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5145..5162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5316..5904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5396..5415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6075..6866,6867..6935))
                     /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     6698..6717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6936..7040)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7108..7126
                     /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 acctcgcgaa
      421 tgcatctaga tctttgtata gaaaagttgg cacctttggt cttttattgt caacttccat
      481 tggttcttcc attgtttctg ttaaattaat gaatttttca taaaataaag acattataca
      541 atataaaaat gaagaattta ttgaaaataa actgccagag agaaaaagta tgcaacactc
      601 ccgccgagag tgtttgaaat ggtgtacggt acattttcgt gctaggagtt agatgtgcag
      661 gcagcaacga gagggggaga gatttttttg ggccttgtga aattaacgtg agttttctgg
      721 tcatctgact aatcatgttg gttttttgtt ggtttatttt gtttttatct ttgtttttat
      781 ccagattagg aaatttaaat tttatgaatt tataatgagg tcaaacattc agtcccagcg
      841 tttttcctgt tctcactgtt tagtcgaatt tttattttag gctttcaaca aatgttctaa
      901 ctgtcttatt tgtgacctca ctttttatat ttttttaatt tttaaaaata ttagaagttt
      961 ctaggataat tttttcgact tttattctct ctaccgtccg cactcttctt acttttaaat
     1021 taaattgttt ttttttcagt tgggaaacac tttgctcaca agtttgtaca aaaaagcagg
     1081 ctccatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
     1141 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
     1201 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catgggtaag
     1261 tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa cacttgtcac
     1321 tactttctgt tatggtgttc aatgcttctc gagataccca gatcatatga aacggcatga
     1381 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
     1441 tgacgggaac tacaagacac gtaagtttaa acagttcggt actaactaac catacatatt
     1501 taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
     1561 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
     1621 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttgtaag
     1681 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact tcaaaattag
     1741 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     1801 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     1861 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     1921 gattacacat ggcatggatg aactatacaa atgcccgggg gatcggtgga gctccaccgg
     1981 tggcggccgc tctagaacta gtccaccaaa gccatctgcc aagggagcca agaaggccgc
     2041 caagaccgtc gttgccaagc caaaggacgg aaagaagaga cgtcatgccc gcaaggaatc
     2101 gtactccgtc tacatctacc gtgttctcaa gcaagttcac ccagacaccg gagtctcctc
     2161 caaggccatg tctatcatga actccttcgt caacgatgta ttcgaacgca tcgcttcgga
     2221 agcttcccgt cttgctcatt acaacaaacg ctcaacgatc tcatcccgcg aaattcaaac
     2281 cgctgtccgt ttgattctcc caggagaact tgccaagcac gccgtgtctg agggaaccaa
     2341 ggccgtcacc aagtacactt ccagcaagta aacccagctt tcttgtacaa agtgggataa
     2401 atgcaaaatc ctttcaagca ttcccttctt ctctatcact cttctttctt tttgtcaaaa
     2461 aattctctcg ctaatttatt tgctttttta atgttattat tttatgactt tttatagtca
     2521 ctgaaaagtt tgcatctgag tgaagtgaat gctatcaaaa tgtgattctg tctgatgtac
     2581 tttcacaatc tctcttcaat tccattttga agtgctttaa acccgaaagg ttgagaaaaa
     2641 tgcgagcgct caaatatttg tattgtgttc gttgagtgac ccaacaaaaa gaggaaactt
     2701 tattgtgccg ccaagaaaaa agtctcacaa ctatcgccta gttctagaca ttctctaatg
     2761 aaaaaatctt tcagttgaaa ttgaaaatga gttaaagttg gagtttttat tgaaaacaga
     2821 tttccgtgtg attagtgttt ttagcgagtg tgacaggaca gcgaaaaaat atagaaacaa
     2881 ggggggaact gaaaagctta ggaatgcatt gaacatgaga aggggaaggg gaaggaacaa
     2941 actagacagg aattattgga atttaatcac atttggagtt ttttttctat tcgacagaat
     3001 aattatccag aacatttttg tattaaatat ttatgcatca tatgagtagt cggctttgtt
     3061 gtgcatgacg agtttgttat cgacgaaata gaagctgtca gaacgagtct cgtttggatt
     3121 gttgatcatg tcgtccactg aaaaagagat tagtcttttg aattgtactt tttagaataa
     3181 tgactcactg agttgttgag agagttgagg gaactcatag atatgttcac agttgtttcg
     3241 tgaattcgga atacagaatc cgaattcaaa gtcaaaacac ttcagaaggc gatctttgaa
     3301 gaagtgacgt tcgatcattc ggaaatgatg gattgggatg tctcctactt tgaattccac
     3361 agttgctccg accgttttga gtttcaaaaa gtttggagca aatctataac gcacgtatct
     3421 tgccgactcc tgtggcgact catcattctc ctgatcgttc tccggtttgg cgatctcgaa
     3481 aagcacttgc tcagtgtcca gatcacggat ttggaacttg gtgaactcga tgttatagat
     3541 gttcgcagat ggggagcata agaatcctaa atttatgttt taaactgaaa tccaaaggga
     3601 gcaagatacc ttgagtgatt cccggaagtg ctaaaacgtc gttcggagtg atttgagctt
     3661 tcttcgcaag ctccgattcc gttgtgattc cttgttcggt gcttggtggt ggccgtggca
     3721 tctggaaata tggaaaagtt caacaaaaag aaaagagaaa agaatgaaat cggatatcaa
     3781 gagttagttg agcggtttct ctagttttct gagtctcacc tgcgacggga aggtcgccga
     3841 gccgggtgga atcgattgtt gttgctcggc tttcatatcg gtttggttgg aagcggctga
     3901 aaacggaaag aagtggaaga aggaaaagag tgtggtgtga caggaaaatg gtaattagag
     3961 ggtgccaaat aaccagctat attttgtttt tttttgaaaa catttttaaa aagaaaaata
     4021 cgataatgat atcagatgga tttccggaaa actggtatga aaaatttcaa cctttttgag
     4081 tacatgtaat caaaatacac tttgtaaatt atcattttta ttgaaactcc accatttttc
     4141 tatttataac gctaataatt tgaaaaagaa acctattgcg aaccgcgggg tgaatcccaa
     4201 aaacgaatgc gttttggtgg agtgattgat tcgaatcgaa gaagaaaaag aagaagacgt
     4261 ggaatagaga gctcactctt aaccgagcag cacacaccga cagaaaaaaa aatgaaatga
     4321 atgagggtct tcttcttctt cttcttcgaa tgattgacag aaatgggaaa aagaggaaga
     4381 ttgagaaggg aaaaaggaag gagaaaagaa gcagaagaag acgtcagaga ggagaggaac
     4441 gagcggaaaa gcagcgggcg caagtcatag aagtagcaga gctggggaga agaagacact
     4501 atccaagaaa ggaatgacga gagagtatgc aaaggggtat agggtgcaga cagaatagga
     4561 acagaataac agatgatgag ccaagaagag ttgaaaaggg cgatgaattt gtcatgtaac
     4621 ttaatttggg tcaatttgag catgatgaat tgaaatcatc ccttgttggg agttaataac
     4681 cggtttgtta tcagaaaccc tgtaatagaa gggcgcccta actttgagcc aattcatccc
     4741 ggtttctgtc aaatatatca aaaagtggtc aactgacaaa ttgtttttga tattataata
     4801 aacattttat ccgttaacaa ttttcgaata ctttttacaa ggacttggat aaattggctc
     4861 aaagccgggc ccgtcgactg cagaggcctg catgcaagct tggcgtaatc atggtcatag
     4921 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4981 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5041 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5101 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6121 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6181 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6241 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6301 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6361 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6421 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6481 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6541 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6601 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6661 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6721 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6781 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6841 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6901 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7081 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
