LOCUS Exported 3596 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION In situ hybridization and RNAi studies in planarians. ACCESSION . VERSION . KEYWORDS Pmo_frz-4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3596) AUTHORS Accorsi A, Williams MM, Ross EJ, Robb SMC, Elliott SA, Tu KC, Sanchez Alvarado A TITLE Hands-On Classroom Activities for Exploring Regeneration and Stem Cell Biology with Planarians JOURNAL The American Biology Teacher 79(3):208-223. Mar 2017. REFERENCE 2 (bases 1 to 3596) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3596 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator 24..71 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 76..94 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 152..168 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 152..168 /label=pBluescriptSK /note="For pBluescript vector" promoter complement(1226..1244) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(1302..1320) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1325..1341) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1325..1341) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1443..1462) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1454..2263 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1521..1540) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2410..2998 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2899..2918 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3152..3169 /label=L4440 /note="L4440 vector, forward primer" terminator 3328..3355 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3447..3533 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 3582..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3596..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtctt aagcaaaaaa cccctcaaga cccgtttaga ggccccaagg 61 ggttatgcta ggaagtaata cgactcacta tagggagacc ggcagatctg atatcatcga 121 tgaattcgag ctccaccgcg gtggcggccg ctctagaact agtggatcca ccggttccat 181 ggctagcggc attaccatcc cggcagttgg actttttagt gggcaacggc aacgagacga 241 cctggtgtaa gattaatttc ttgatgctct actttttcag ttcagccagc gcgttgtggt 301 gggtcgtttt aacagtcacg tggtttcttt cggcatctcg gcattggggg tatgaggcta 361 ttgaatcagt cagcagtatt ttgcatctag tcagttgggg aattccggcg ctcaagtcaa 421 ttttcgttct catccttcac aaaatcgacg ccgatgaatt aactggacaa tgttacgtcg 481 gcaactcaga taaggaagcg ctgattggtt tcattctagt cccacaagtt ctctacttag 541 tgaccggggt cacgttccta gcgttgggct actactccat cttgaaggta agaaagtctt 601 tcctacaaca ggtgagttgt attgacacca cgaatttgaa acgacttgaa aaattaatgg 661 cgaaaatcgg cgtcttttcc attttatata ttgtaccagt cacgtgtaca attataagct 721 acgttatcga catcttcagg acttcctatt tcgatttaat gctgaaggcg ttgtatgaat 781 attcagcaga ttgtgtcaca gtaaagggtc tgtcgtggtc gttggcgaag tgcgtagaca 841 tcattcaacc ggcattgccg tccatcgaag tgaggatgct gcgtattttt atgaatcttg 901 tcatcggcat aacatcgggc atttggattt ggggtaacaa aaagtcgttc cgatattgtg 961 ggcgaaataa aaaagaaatt caacaatgtg aaatgagaca aatcgaagta ttaacccccc 1021 tcaatcagac tccgtttatt aaatccggtg aggacacttt tccccatcac gggtggagtt 1081 atgtgccaac cttggcctcg gaacacgcgg tcggggggct ctctcagacc cctgtggaag 1141 gtataatggc ccccaataac tcatcaataa gcgaagtatc cagtaagcat cgggtagaat 1201 tggccgggcc cggtacccaa ttcgccctat agtgagtcgt attacttcca cataacccct 1261 tggggcctct aaacgggtct tgaggggttt tttgaggatc ccctatagtg agtcgtatta 1321 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat 1381 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa 1441 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa 1501 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc 1561 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa 1621 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt 1681 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac 1741 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga 1801 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa 1861 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa 1921 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt 1981 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga 2041 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg 2101 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga 2161 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat 2221 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg 2281 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa 2341 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa 2401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 2461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 2521 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 2581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 2641 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 2701 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 2761 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 2821 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 2881 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 2941 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3001 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 3061 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 3121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 3181 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 3241 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 3301 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 3361 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 3421 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 3481 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 3541 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgtt //