LOCUS Exported 4084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Provides the CDS of the Hygromycin resistance gene as a level 0 GoldenBraid part. ACCESSION . VERSION . KEYWORDS pHygromycin (GB0211) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4084) AUTHORS Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D TITLE GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. JOURNAL Plant Physiol. 2013 Jul;162(3):1618-31. PUBMED 23669743 REFERENCE 2 (bases 1 to 4084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4084 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 6..1031 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" promoter complement(1134..1152) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1135..1152) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1170..1186) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1170..1186) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1183..1205) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1194..1210 /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(1218..1248) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1263..1284 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1401..1418) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1572..2160) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1652..1671) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2331..3191) /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 2954..2973 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3192..3296) /gene="bla" /label=AmpR promoter rep_origin complement(3374..3829) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3511..3532) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3723..3742 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3955..3977 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3969..3986 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3970..3986 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3993..4012 /label=T7 /note="T7 promoter, forward primer" promoter 3993..4011 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 4042..4051 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 4046..4055 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 ctcgaatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt 61 tcgacagcgt gtccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct 121 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca 181 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg 241 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca 301 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca 361 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc 421 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg 481 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg 541 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt 601 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg 661 aggcaatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt 721 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat 781 cgccacgact ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg 841 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat 901 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg 961 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg 1021 caaaggaatg agcttcgaga ccactcatcg cccaatcact agtgaattcg cggccgcctg 1081 caggtcgacc atatgggaga gctcccaacg cgttggatgc atagcttgag tattctatag 1141 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 1201 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 1261 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 1321 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 1381 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 1441 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 1501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 1561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 1621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 1681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 1741 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 1801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 1861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 1921 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 1981 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 2041 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 2101 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 2161 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 2221 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 2281 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 2341 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 2401 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 2461 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 2521 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 2581 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 2641 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 2701 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 2761 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 2821 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 2881 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 2941 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 3001 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 3061 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 3121 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 3181 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 3241 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 3301 tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa 3361 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt 3421 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 3481 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 3541 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 3601 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 3661 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 3721 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 3781 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat 3841 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc 3901 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt 3961 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg 4021 gcccgacgtc gcatgctccc ggccgccatg gcggccgcgg gaattcgatt gggcgatgag 4081 tggt //