LOCUS       sequence-111542-        5011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses gfp::h2b in C. elegans cells from the ubiquitous eef-1A.1 
            (eft-3) promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5011)
  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 5011)
  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: pIK127
FEATURES             Location/Qualifiers
     source          1..5011
                     /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"
     primer_bind     complement(2790..2806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2790..2806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2803..2825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2814..2830
                     /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(2838..2844,2845..2862,2863..2868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2883..2904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3021..3038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3192..3780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3272..3291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3951..4742,4743..4811))
                     /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     4574..4593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4812..4916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4984..5002
                     /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 ctatcggatc ccgggcccgt cgactgcaga
     2761 ggcctgcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2821 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2881 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2941 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3001 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3061 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4081 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4921 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4981 aaaaataggc gtatcacgag gccctttcgt c
//
