LOCUS Exported 2752 bp ds-DNA circular SYN 05-DEC-2018 DEFINITION MoClo golden gate assembly CD part for RhlI (quorum-sensing synthase for 3OC14).. ACCESSION . VERSION . KEYWORDS C36m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2752) TITLE CIDAR MoClo Extension REFERENCE 2 (bases 1 to 2752) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 5, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers ligation 1..4 /label="Ligation" CDS 2..604 /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /Transferred_From="C165_pLLacBCD2ms (modified), C216_pRhl17LacOs (modified), C247_pRhlo17LacO (modified)" /Transferred_Similarity="100.00%" /label="RhlI from PAO1" misc_feature 5..24 /Mismatches=0 /Extension="TAGTGGAAGACGTAATG" /created_by="primer3" /annotation_group="281738780596554a8ba690a-5d0d-4004-8922- ea495bdcafd5" /%GC="40.0" /Tm="53.2" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Sequence="ATCGAATTGCTCTCTGAATC" /label="pr41a GG RhlI Fwd ohC" /note="Geneious type: primer_bind_reverse" misc_feature 74..248 /Source="referenceToDocument: urn:local:.:6s-9u0niyb" /label="Digestion of RhlI - Fragment 3 (concatenated sequence 2)" /note="Geneious type: Concatenated sequence" ligation 74..77 /label="Ligation" misc_feature complement(80..85) /Recognition_pattern="GAAGAC(2/6)" /annotation_group="GAAGAC" /label="BbsI" /note="Geneious type: restriction site" misc_feature 109..114 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 137..142 /Recognition_pattern="G^AATTC" /annotation_group="GAATTC" /label="EcoRI" /note="Geneious type: restriction site" misc_feature 245..652 /Source="referenceToDocument: urn:local:.:6s-9u0niyb" /label="Digestion of RhlI - Fragment 4 (concatenated sequence 3)" /note="Geneious type: Concatenated sequence" ligation 245..248 /label="Ligation" misc_feature complement(251..256) /Recognition_pattern="GAAGAC(2/6)" /annotation_group="GAAGAC" /label="BbsI" /note="Geneious type: restriction site" misc_feature complement(497..502) /Recognition_pattern="CGTCTC(1/5)" /annotation_group="CGTCTC" /label="BsmBI" /note="Geneious type: restriction site" sig_peptide 611..643 /ApEinfo_fwdcolor="#ff8000" /ApEinfo_revcolor="#ff8000" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /ApEinfo_label="ssrA" /Transferred_From="C165_pLLacBCD2ms (modified), C205a_pRhlLacsfC (modified), C216_pRhl17LacOs (modified), C217_pCino17LacO (modified), C235_pCino17tetm (modified), C247_pRhlo17LacO (modified)" /Transferred_Similarity="100.00%" /label="ssrA; ssrA(1)" misc_feature 649..77 /label="Digestion of DVA_CD (modified) - Fragment 2 (concatenated sequence 1)" /note="Geneious type: Concatenated sequence" ligation 649..652 /label="Ligation" sig_peptide 653..2752 /ApEinfo_revcolor="#6ecff6" /ApEinfo_fwdcolor="#6ecff6" /label="DVLA" misc_feature complement(654..659) /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 661..666 /Recognition_pattern="A^CTAGT" /annotation_group="ACTAGT" /label="SpeI" /note="Geneious type: restriction site" misc_feature 2739..2744 /Recognition_pattern="A^CTAGT" /annotation_group="ACTAGT" /label="SpeI" /note="Geneious type: restriction site" misc_feature 2746..2751 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" misc_feature 2746..2751 /Recognition_pattern="GGTCTC(1/5)" /annotation_group="GGTCTC" /label="BsaI" /note="Geneious type: restriction site" source 1..2752 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 660..680 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(686..703) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(857..1445) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(937..956) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1616..2476) /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 2239..2258 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2477..2581) /gene="bla" /label=AmpR promoter primer_bind 2649..2667 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2671..2714 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 2717..2738 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 aatgatcgaa ttgctctctg aatcgctgga agggctttcc gccgccatga tcgccgagct 61 gggacgctac cggcatcagg tcttcatcga gaagctgggc tgggacgtgg tctccacctc 121 cagggtccgc gaccaggaat tcgaccagtt cgaccatccg caaacccgct acatcgtcgc 181 catgagccgc cagggcatct gcggttgcgc ccgcctgctg ccgacgaccg acgcctacct 241 gctcaaggac gtcttcgcct acctgtgcag cgaaaccccg ccgagcgatc cgtcggtctg 301 ggagctttcg cgctacgccg ccagcgcggc ggacgatccg cagctggcga tgaagatatt 361 ctggtccagc ctgcaatgcg cctggtacct gggcgccagt tcggtggtgg cggtgaccac 421 cacggccatg gagcgctatt tcgttcgcaa cggcgtgatc ctccagcgcc tcggcccgcc 481 gcagaaggtc aagggcgaga cgctggtcgc gatcagcttc ccggcctacc aggagcgcgg 541 cctggagatg ctgctgcgct accacccgga atggctgcag ggcgtaccgc tgtcgatggc 601 ggtgacctcc gcagcgaacg acgaaaatta cgcccttgca gcgtaataag gtagagacct 661 actagtagcg gccgctgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 841 ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac aaaaatcgac 901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 1021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 1081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 1141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 1201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 1261 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 1321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 1381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 1441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 1501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 1561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 1621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 1681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 1741 ctgcaatgat accgcgcgac ccacgctcac cggctccaga tttatcagca ataaaccagc 1801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 1861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 1921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 1981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 2041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 2101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 2161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 2221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 2281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 2341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 2401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 2461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 2521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 2581 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa 2641 cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg 2701 ctaaggatga tttctggaat tcgcggccgc ttctagagac tagtgggtct ca //