LOCUS Exported 3078 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo golden gate assembly AE destination vector DVCK-AE-GFP (ColE1/kan resistance destination vector with constitutive GFP expression for screening.. ACCESSION . VERSION . KEYWORDS V35m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3078) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 3078) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3078 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 6..24 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 28..71 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 74..95 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" CDS 224..940 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(254..282) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 877..897 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 956..1027 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1043..1070 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 1180..1200 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1201..1258 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined." primer_bind complement(1282..1299) /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(1507..1526) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1518..2333 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1591..1610) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin complement(2455..3043) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2535..2554) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gcataaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta 61 aggatgattt ctggaattcg cggccgcttc tagagactag tggaagacat ggagagagac 121 ctgcaccata tgcggtgaga ctagtggaag acatggagtt gacagctagc tcagtcctag 181 gtactgtgct agctactaga gtcacacagg aaagtactaa atgatgcgta aaggagaaga 241 acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa 301 attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt 361 tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta ctttcggtta 421 tggtgttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact ttttcaagag 481 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta 541 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa 601 aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa 661 ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa 721 aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac 781 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc 841 cctttcgaaa gatcccaacg aaaagagaga tcacatggtc cttcttgagt ttgtaacagc 901 tgctgggatt acacatggca tggatgaact atacaaataa taaaggtcca ggcatcaaat 961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa 1021 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatagctt 1081 atgtcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 1141 tgaatcggcc aacgcgcggg ggtctctgct tatgtcttct actagtagcg gccgctgcag 1201 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt 1261 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 1321 gcgagcggta tcagctcact caaaggcggt aagtcgcgtg ttatacgccc gttatccatg 1381 ggtatggaca gttttccctt tgatatgtaa cgcacgttgt gtctcaaaat ctctgatgtt 1441 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca 1501 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctcc cgtccgcgct 1561 taaactccaa catggacgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc 1621 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga 1681 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtccgt ctcaactggc 1741 tgacggagtt tatgcctctc ccgaccatca agcattttat ccgtactcct gatgatgcgt 1801 ggttactcac caccgcgatt cctgggaaaa cagccttcca ggtattagaa gaatatcctg 1861 attcaggtga aaatattgtt gatgcgctgg ccgtgttcct gcgccggtta cattcgattc 1921 ctgtttgtaa ttgtcctttt aacagcgatc gtgtatttcg tcttgctcag gcgcaatcac 1981 gcatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg 2041 ttgaacaagt ctggaaagaa atgcacaagc tcttgccatt ctcaccggat tcagtcgtca 2101 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta 2161 ttgatgttgg acgggtcgga atcgcagacc gttaccagga ccttgccatt ctttggaact 2221 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata 2281 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taataatact 2341 agcagaaatc atccttagcg aaagctaagg atttttttta tctgattacc gcctttgagt 2401 gagcgtcgac ctagtcctag ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 2461 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 2521 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 2581 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 2641 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 2701 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 2761 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 2821 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 2881 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 2941 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 3001 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 3061 ttctacgggg tctgacgc //