LOCUS Exported 3092 bp ds-DNA circular SYN 05-DEC-2018 DEFINITION MoClo golden gate assembly CD part treRT-LAA (synthetic Lac-family repressor chimera with LAA degradation tag for ssrA; see DOI: 10.1021/sb500262f).. ACCESSION . VERSION . KEYWORDS C102m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3092) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 3092) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 5, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers misc_feature 3074..966 /Source="referenceToDocument: urn:local:.:ei-9u0ntrr" /label="AS079 PCR product treRT with overhangs (concatenated sequence 1)" /note="Geneious type: Concatenated sequence" CDS 5..946 /created_by="User" /modified_by="anandhswaminathan" /Transferred_From="pZS1 (treRT,fruRT) - treRT" /Transferred_Similarity="100.00%" /label="treRT" misc_feature 139 /ApEinfo_fwdcolor="#ff0000" /ApEinfo_revcolor="#ff0000" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /label="G -> C" misc_feature complement(934..946) /Mismatches=0 /Extension="TCGTCGTTCGCTGCACTAGT" /%GC="76.9" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Pair_Dimer_Tm="None" /Tm="51.3" /annotation_group="25606898581648286e47824d-1024-4fd6-8076 -1c0f76a69f9f" /Sequence="GGACAGGGTGGCG" /created_by="primer3" /Product_Size=942 /label="treRT Rev, OH to LAA" /note="Geneious type: primer_bind_reverse" misc_feature <947..3092 /Original_Bases /label /note="Geneious type: Editing History Insertion" misc_feature 947..3092 /Source="referenceToDocument: urn:local:.:eh-9u0ntrr" /label="C20m PCR Product LAA -> DVLA BB (concatenated sequence 2)" /note="Geneious type: Concatenated sequence" ligation 947..966 /label="Ligation" CDS <947..988 /codon_start=1 /created_by="User" /label="sfYFP ssrA" misc_feature 947..988 /ApEinfo_fwdcolor="#ff8040" /ApEinfo_revcolor="#ff8040" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /label="LAA" misc_feature 993..3092 /ApEinfo_revcolor="#6ecff6" /ApEinfo_fwdcolor="#6ecff6" /label="DVLA" misc_feature complement(994..999) /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature complement(1018..1032) /Mismatches=0 /%GC="66.7" /Hairpin_Tm="38.0" /Self_Dimer_Tm="None" /Tm="53.0" /annotation_group="239505150997683517b43e81-82e4-4d0d-b74e -4669b0a43f11" /Sequence="GAGCGAGGAAGCCTG" /created_by="primer3" /label="DVLA universal downstream Rev" /note="Geneious type: primer_bind_reverse" misc_feature complement(1018..1032) /Mismatches=0 /%GC="66.7" /Hairpin_Tm="38.0" /Self_Dimer_Tm="None" /Pair_Dimer_Tm="None" /Tm="53.0" /annotation_group="2395152314946931566eb017-50ea-4b44-9f41 -869fc2aa2930" /Sequence="GAGCGAGGAAGCCTG" /created_by="primer3" /Product_Size=86 /label="DVLA universal downstream Rev" /note="Geneious type: primer_bind_reverse" primer_bind complement(1076..1096) /Sequence="TTACCGCCTTTGAGTGAGCTG" /Hairpin_Tm="None" /Tm="60.6" /Self_Dimer_Tm="None" /%GC="52.4" /created_by="primer3" /label="SC190/Part.Seq.REV" primer_bind 2988..3008 /Sequence="AAATAGGCGTATCACGAGGCA" /Hairpin_Tm="46.5" /Tm="59.6" /Self_Dimer_Tm="None" /%GC="47.6" /created_by="primer3" /label="SC189/Part.Seq.FOR" misc_feature complement(3070..3092) /Mismatches=0 /%GC="47.8" /Hairpin_Tm="54.8" /Self_Dimer_Tm="19.3" /Pair_Dimer_Tm="10.7" /Tm="57.9" /annotation_group="2560630355998674683a42e4-7deb-4c08-84d4 -b42b0ae6dbf6" /Sequence="TGAGACCCACTAGTCTCTAGAAG" /created_by="primer3" /Product_Size=2146 /label="DVLA universal upstream Rev" /note="Geneious type: primer_bind_reverse" ligation 3074..3092 /label="Ligation" misc_feature 3086..3091 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 3086..3091 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" source 1..3092 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(24..43) /label=LacI-R /note="LacI, reverse primer" misc_feature 1000..1020 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(1026..1043) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1197..1785) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1277..1296) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1956..2816) /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 2579..2598 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2817..2921) /gene="bla" /label=AmpR promoter primer_bind 2989..3007 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 3011..3054 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 3057..3078 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 aatgatgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct ctacggcgac 61 cgtttccaac gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga 121 agcggcgatg gcggagctca attacattcc caaccgcgcc gcacaacaac tggcgggcaa 181 acagagcaaa gtggtcgcca tcattgttac ccgtctggat tcgttgtcag aaaatctcgc 241 cgttcaaacc atgctgccag cgttctatga acaaggttac gacccaatca tgatggaaag 301 tcagttttcc ccgcaattag ttgccgaaca tttgggagtg ctgaaacggc gtaatatcga 361 cggcgtagtg ctgttcggtt ttactggcat aacagaagaa atgttagccc actggcagtc 421 atcgctggtt ctgctggcgc gtgacgcaaa aggctttgct tcggtctgtt atgacgacga 481 aggggcaatc aaaatcctga tgcaacggct gtatgaccag gggcatcgta atatcagtta 541 tctcggcgtg ccgcacagtg acgtgacaac cggtaagcga cgtcacgaag cctacctggc 601 gttctgcaaa gcgcataaac tgcatcccgt tgccgccctg ccagggcttg ctatgaagca 661 aggctatgag aacgttgcaa aagtgattac gcctgaaact accgccttac tgtgcgcaac 721 cgacacgctg gcacttggcg caagtaaata cctgcaagag caacgcatcg acaccttgca 781 actggcgagc gtcggtaata cgccgttaat gaaattcctc catccggaga tcgtaaccgt 841 agatcccggt tacgccgaag ctggacgcca ggcggcttgc cagttgatcg cgcaggtaac 901 cgggcgcagc gaaccgcaac aaatcatcat ccccgccacc ctgtccacta gtgcagcgaa 961 cgacgaaaat tacgcccttg cagcgtgaag gtagagacct actagtagcg gccgctgcag 1021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 1081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 1141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 1201 cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 1261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 1321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 1381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 1441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 1501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 1561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 1621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 1681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 1741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 1801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 1861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 1921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 1981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 2041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgcgac 2101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 2161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 2221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 2281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 2341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 2401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 2461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 2521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 2581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 2641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 2701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 2761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 2821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 2881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 2941 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc 3001 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat 3061 tcgcggccgc ttctagagac tagtgggtct ca //