LOCUS       sequence-299633-        4339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (eft-3p::wrmScarlet::tbb-2 3'UTR in pUC19) Experimentally used to 
            mark formation of transgenic arrays for C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4339)
  AUTHORS   Stevenson ZC, Moerdyk-Schauwecker MJ, Jamison B, Phillips PC
  TITLE     Rapid Self-Selecting and Clone-Free Integration of Transgenes into 
            Engineered CRISPR Safe Harbor Locations in Caenorhabditis elegans.
  JOURNAL   G3 (Bethesda). 2020 Aug 19. pii: g3.120.401400. doi: 
            10.1534/g3.120.401400.
  PUBMED    32816924
REFERENCE   2  (bases 1 to 4339)
  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). 2020 Aug 19. pii: g3.120.401400. doi: 
            10.1534/g3.120.401400."
COMMENT     Sequence Label: pZCS16 (Peft-3::wrmScarlet::tbb-2 3'UTR in pUC19)
FEATURES             Location/Qualifiers
     source          1..4339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    956..977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(992..997,998..1015,1016..1022)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1030..1046
                     /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     1035..1057
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1054..1070
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1054..1070
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1120..1719
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     terminator      2446..2769
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2793..2810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2793..2809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2802..2824)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3018..3037)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3137..3159
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3197..3215)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3283..3387
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3388..3456,3457..4248)
                     /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(3606..3625)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      901 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      961 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     1021 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     1081 ccaagcttgc atgcctgcag gtcgactcta gaggatcccg cacctttggt cttttattgt
     1141 caacttccat tggttcttcc attgtttctg ttaaattaat gaatttttca taaaataaag
     1201 acattataca atataaaaat gaagaattta ttgaaaataa actgccagag agaaaaagta
     1261 tgcaacactc ccgccgagag tgtttgaaat ggtgtacggt acattttcgt gctaggagtt
     1321 agatgtgcag gcagcaacga gagggggaga gatttttttg ggccttgtga aattaacgtg
     1381 agttttctgg tcatctgact aatcatgttg gttttttgtt ggtttatttt gtttttatct
     1441 ttgtttttat ccagattagg aaatttaaat tttatgaatt tataatgagg tcaaacattc
     1501 agtcccagcg tttttcctgt tctcactgtt tagtcgaatt tttattttag gctttcaaca
     1561 aatgttctaa ctgtcttatt tgtgacctca ctttttatat ttttttaatt tttaaaaata
     1621 ttagaagttt ctaggataat tttttcgact tttattctct ctaccgtccg cactcttctt
     1681 acttttaaat taaattgttt ttttttcagt tgggaaacac tttgctctag aaaaaatggt
     1741 cagcaaggga gaggcagtta tcaaggagtt catgcgtttc aaggtccaca tggagggatc
     1801 catgaacgga cacgagttcg agatcgaggg agagggagag ggacgtccat acgagggaac
     1861 ccaaaccgcc aagctcaagg tcaccaaggg aggaccactc ccattctcct gggacatcct
     1921 ctccccacaa ttcatgtacg gatcccgtgc cttcaccaag cacccagccg acatcccaga
     1981 ctactacaag caatccttcc cagagggatt caagtgggag cgtgtcatga acttcgagga
     2041 cggaggagcc gtcaccgtca cccaagacac ctccctcgag gacggaaccc tcatctacaa
     2101 ggtcaagctc cgtggaacca acttcccacc agacggacca gtcatgcaaa agaagaccat
     2161 gggatgggag gcctccaccg agcgtctcta cccagaggac ggagtcctca agggagacat
     2221 caagatggcc ctccgtctca aggacggagg acgttacctc gccgacttca agaccaccta
     2281 caaggccaag aagccagtcc aaatgccagg agcctacaac gtcgaccgta agctcgacat
     2341 cacctcccac aacgaggact acaccgtcgt cgagcaatac gagcgttccg agggacgtca
     2401 ctccaccgga ggaatggacg agctctacaa gtaaactagt gataaatgca aaatcctttc
     2461 aagcattccc ttcttctcta tcactcttct ttctttttgt caaaaaattc tctcgctaat
     2521 ttatttgctt ttttaatgtt attattttat gactttttat agtcactgaa aagtttgcat
     2581 ctgagtgaag tgaatgctat caaaatgtga ttctgtctga tgtactttca caatctctct
     2641 tcaattccat tttgaagtgc tttaaacccg aaaggttgag aaaaatgcga gcgctcaaat
     2701 atttgtattg tgttcgttga gtgacccaac aaaaagagga aactttattg tgccgccaag
     2761 aaaaaagtct cacgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac
     2821 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2881 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2941 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3001 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3061 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3121 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3181 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3241 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3301 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3361 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3421 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3481 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3541 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3601 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3661 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3721 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3781 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3841 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3901 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3961 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4021 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4081 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4141 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4201 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4261 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4321 gtgaagatcc tttttgata
//
