LOCUS       sequence-213065-        5976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High affinity Myc tagging of weakly expressed genes (optimized for 
            Toxoplasma gondii).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5976)
  AUTHORS   Hortua Triana MA, Marquez-Nogueras KM, Chang L, Stasic AJ, Li C, 
            Spiegel KA, Sharma A, Li ZH, Moreno SNJ
  TITLE     Tagging of Weakly Expressed Toxoplasma gondii Calcium-Related Genes 
            with High-Affinity Tags.
  JOURNAL   J Eukaryot Microbiol. 2018 Apr 19. doi: 10.1111/jeu.12626.
  PUBMED    29672999
REFERENCE   2  (bases 1 to 5976)
  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: "J Eukaryot 
            Microbiol. 2018 Apr 19. doi: 10.1111/jeu.12626."
COMMENT     Sequence Label: pLIC-SM-Myc
FEATURES             Location/Qualifiers
     source          1..5976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             338..367
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             371..400
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             404..433
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             965..994
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1001..1030
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1043..1072
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1079..1108
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1322..1351
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1355..1384
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1388..1417
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1846..1862
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1847..1863
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2378..3040
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MHEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
                     LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS
                     SLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN
                     MDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQVGA"
     primer_bind     complement(2447..2466)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3358..3374)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3406..3425)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3407..3425)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3432..3449)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3432..3448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3441..3463)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3590..4045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3677..3696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3887..3908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4071..4175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4176..4244,4245..5036)
                     /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(4394..4413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5207..5795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5696..5715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5949..5966
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
      241 aaattaaccc tcactaaagg gaacaaaagc tggtaccctg tacttccaat ccaatttaat
      301 taaaattgga agtggaggac gggaattccc taggatggag caaaaactga tatcggagga
      361 agacttggca gagcagaaac tgatttcgga agaggatctt gccgaacaaa aactgatcag
      421 tgaggaagac cttgccggag tgtcgaaggg tgaagagctg ttcaccggcg tggtgcccat
      481 cctcgtcgag ctggacggag atgtcaatgg acataaattc tccgtacggg gagagggcga
      541 gggcgacgca accaacggta aactgaccct gaagttcatt tgcaccacgg gtaaactgcc
      601 cgtaccctgg cccaccctgg tcacaacctt gggtggcgga gttcagtgtt tttcgcggta
      661 tccagatcac atgaagcagc atgacttttt caagagcgct atgcccgagg gttacgttca
      721 agagaggacc atcagtttca aggacgacgg cacctacaag acacgggctg aggtgaaatt
      781 cgagggtgat acactcgtga atcgtatcga gctgaagggc atagatttta aggaggatgg
      841 caatatcctg ggccacaagt tggagtataa cttcaacagt cacaatgtct atataactgc
      901 cgataagcag aagaatggca ttaaagccaa tttcaaaatt cgtcacaatg tcgaaggcgg
      961 cgcagaacag aagctcatca gtgaagagga tctggccgca gagcagaaac tcattagcga
     1021 ggaggacttg ggcggaggcg gcgagcagaa gctgatatcg gaggaagacc tggcggccga
     1081 gcaaaagctc atctcggagg aggatctcgc gggaggagat ggaagtgtcc agttggcgga
     1141 tcactaccag cagaatacgc caattggtga cggccccgtc cttctgccgg ataaccacta
     1201 cctctccacc cagtcggtgc tgagcaagga cccgaatgaa aaacgtgacc acatggttct
     1261 tcttgagttc gtcacggctg cgggcatcac gttgggcatg gacgaactgt acaagggcgc
     1321 tgaacagaaa cttatctccg aggaggatct ggcggaacag aagctgatca gcgaagagga
     1381 cctggcggag cagaaactta tcagcgaaga ggatctgtaa catatgtaga aaagttgtaa
     1441 cgttagtaaa cgtaacacac gggtataata gccatttaat ggcgccgtat atttttcact
     1501 ccccctctcc gataggagga agaaagacga tccgtctgcg aatgtatatt acatcgaccg
     1561 gccatagtac gagaagtggt gccggtattg ttctgtccgg ggggtgtacc atcctatttt
     1621 ttgtatatgt cgttcttgtc ggggagcaac agcccatctt ccggctattg tctcgtatca
     1681 gttgtgaaac agactgcgtt ttgccgaaaa tgagtcctat gtccgatggc tatttgtacc
     1741 gcttcgtacc atttcgcttg tcagctgtaa tcaaccgaat tcatttggcg agacatgatt
     1801 ccgattaaat tctttactgc gcgtgctcgg atccactagt tctactcgag gtcgacggta
     1861 tcgataagct tgatggcgat gcatgtcccg cgttcgtgaa attctctgca tcagcggagt
     1921 gatcaggaat catcgtctca gcgggatgac gttgcggagc aggccggctc gcggtgggca
     1981 gtcagatgcc gaaggcgtaa ctcaggacgg cttgcgctca tcgcagaaca ggggtggtgc
     2041 ctgcattggg tgcggttggt gatcctggtt ggaccggtgg agatgcgcgc gcacgaaggg
     2101 gatgtgtcag aaacattttg tttgttctct gtgaactttt agatgtgtta aaggcggcga
     2161 atattagcag agagtcctcc ttgttggatt ctctcttgaa tttcgccctt tccttctctt
     2221 tgcgagtctc gtagagaaca agcactcgtt cgccgtccct gacgacgcaa cccgcgcaga
     2281 agacatccac caaacggtgt tacacaatca ccttgtgtga agttcttgcg gaaaactact
     2341 cgttggcatt ttttcttgaa ttcccttttt cgacaaaatg catgagaaaa aaatcactgg
     2401 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     2461 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     2521 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     2581 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     2641 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     2701 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     2761 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     2821 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2881 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2941 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     3001 attacaacag tactgcgatg agtggcaggt cggggcgtaa ttaatcaccg ttgtgctcac
     3061 ttctcaaatc gacaaaggaa acacacttcg tgcagcatgt gccccattat aaagaaactg
     3121 agttgttccg ttgtggcttg caggtgtcac atccacaaaa accggccgac tctaaatagg
     3181 agtgtttcgc agcaagcagc gaaagtttat gactgggtcc gaatctctga acggatgtgt
     3241 ggcggacctg gctgatgttg atcgccgtcg acacacgcgc cacatgggtc aatacacaag
     3301 acagctatca gttgttttag tcgaaccggt taacacaatt cttgcccccc cgagggggat
     3361 ccactagttc tagagcggcc gccaccgcgg tggagctcca attcgcccta tagtgagtcg
     3421 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3481 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3541 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3601 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3661 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3721 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3781 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3841 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3901 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3961 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4021 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4141 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4201 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4261 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4321 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4381 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4441 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4501 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4561 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4621 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4681 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4741 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4861 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5041 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5341 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5701 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5761 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5821 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5881 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5941 ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
