LOCUS       pFNC-3                  6862 bp ds-DNA     circular     22-NOV-2024
DEFINITION  .
ACCESSION   
VERSION     
KEYWORDS    "Responsible person/group" "Usage" "Status" "Parent plasmids"
            "Cloning sites" "Antibiotic resistance" "Origin of replication"
            "Promoters" "RBS" "Genes" "Tags" "Markers" "Spacers" "gRNAs"
            "Terminators" "DNA fragments" "Protein" "Backbones" "Design entry"
            "LabCollector ID" "Comment" "Box" "Description" "Name"
SOURCE      
  ORGANISM  
FEATURES             Location/Qualifiers
     primer          complement(503..533)
                     /label="Pri_CDV_1163_SacB-gadget-U-rev"
                     /note="sequence: ATTTAAAtGCAAGACCTAAAATGTGTAAAGG"
     primer          526..553
                     /label="FF170_SEVA_AbR_exc_fw"
                     /note="sequence: atttaaaTtggcgaaaatgagacgttga"
     source          530..1321
                     /label="source"
     CDS             654..1313
                     /label="cat CDS"
     gene            654..1313
                     /label="cat gene"
     primer          complement(1300..1325)
                     /label="FF171_SEVA_AbR_exc_rv"
                     /note="sequence: accgaagTcaaattacgccccgccct"
     primer          1318..1343
                     /label="FF168_SEVA_AbR_BB_USER_fw"
                     /note="sequence: acttcggTccgcgcgttgtccttttc"
     source          1322..529
                     /label="from pS6213-bxbI"
     primer          complement(1335..1356)
                     /label="Primer068_AbR-BoxI-rev"
                     /note="sequence: GGGTTATGCAGCGGAAAAGGAC"
     misc_feature    1338..1583
                     /label="oriT"
     primer          complement(1370..1399)
                     /label="Primer072_AbR-USER-rev"
                     /note="sequence: AGGATGGATATACCGAAAAAATCGCTATAA"
     primer          complement(1386..1413)
                     /label="Primer143_pSEVA-OriT-rev"
                     /note="sequence: ATATCGTGCGAAAAAGGATGGATATACC"
     primer          1391..1418
                     /label="Primer069_pSEVA-forAbR-USER-for"
                     /note="sequence: ATCCATCCTTTTTCGCACGATATACAGG"
     primer          1407..1431
                     /label="Primer146_pSEVA-OriT-USER-for"
                     /note="sequence: ACGATATACAGGATTTTGCCAAAGG"
     gene            complement(1642..2790)
                     /label="trfA gene"
     CDS             complement(1642..2790)
                     /label="trfA CDS"
     CDS             complement(1642..2790)
                     /label="Translation 2790-1642"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGSIVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNGDLVHCKR*"
     primer          complement(2767..2790)
                     /label="PM_Promotor_rv"
                     /note="sequence: aaggcctaccccttaggCTTTATGCAAGCTTAGGAGGAAAAACATatgaatcggacgtttgaccggaag"
     primer          3172..3199
                     /label="Primer350_pFREEBB_rv"
                     /note="sequence: AGGCCCGCGTTAGCGGGCCGGGAGGGTT"
     primer          3783..3805
                     /label="NK0459"
                     /note="sequence: GATCTCAAGAAGATCCTTTGATC"
     primer          complement(3805..3830)
                     /label="FF220_pS62xSac_BB_rv"
                     /note="sequence: agacagcTgggcgcgccgtagaaaag"
     primer          3823..3848
                     /label="FF221_pSxx13_fw"
                     /note="sequence: agctgtcTagggcggcggatttgtcc"
     misc_feature    3828..3845
                     /label=2821
     terminator      3828..3932
                     /label="T1"
     primer          3831..3848
                     /label="Pri_CDV_108_pS1"
                     /note="sequence: AGGGCGGCGGATTTGTCC"
     promoter        3943..4007
                     /label="Pem7 promoter"
     primer          complement(3980..4010)
                     /label="FF248_pEM7_U_rv"
                     /note="sequence: attggtTtagttcctcaccttgtcgtattat"
     misc_feature    4028..5530
                     /label="Integrase"
     CDS             4028..5530
                     /label="BxbI"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLDVSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLVVSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGYLPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQGREPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAELVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTVAMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAGSPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSMNVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS*"
     primer          4031..4056
                     /label="FF223_Bxb1_fw"
                     /note="sequence: agagcccTggtagtcatccgcctgtc"
     primer          complement(5511..5537)
                     /label="FF224_Bxb1_rv"
                     /note="sequence: agcgttatTacgacatcccggtgtgta"
     primer          5529..5561
                     /label="FF219_pS62xSac_BB_fw"
                     /note="sequence: aataacgcTgatagtgctagtgtagatcgctac"
     primer          complement(5537..5561)
                     /label="RFP_rv"
                     /note="sequence: AGGACTAGTgtagcgatctacactagcactatca"
     primer          complement(5537..5570)
                     /label="Primer362_RFP-fw"
                     /note="sequence: AGGACTAGTGTAGCGATCTACACTAGCACTATCA"
     primer          5562..5592
                     /label="Primer343_pSEVABB_fw"
                     /note="sequence: ACTAGTCCTAGAGTCGACCTGCAGGCATGCA"
     primer          5562..5592
                     /label="Pri_CDV_376_pTn7-trfA.SalI-for"
                     /note="sequence: ACTAGTCCTAGAGTCGACCTGCAGGCATGCA"
     primer          5562..5592
                     /label="Primer376_pTn7-trfA.SalI-for"
                     /note="sequence: ACTAGTCCTAGAGTCGACCTGCAGGCATGCA"
     primer          complement(5570..5596)
                     /label="PPKp-II-R"
                     /note="sequence: AGCTTGCATGCCTGCAGGTCGACTCTA"
     primer          5573..5604
                     /label="Primer468_pSEVA-RFP-U-for"
                     /note="sequence: AGTCGACCTGCAGGCATGCAAGCTTGCGGCCG"
     primer          5588..5608
                     /label="Primer040_pSEVA-U-for"
                     /note="sequence: ATGCAAGCTTGCGGCCGCGTC"
     primer_bind     complement(5606..5629)
                     /label="F24"
     primer          complement(5606..5629)
                     /label="23-F24"
                     /note="sequence: CGCCAGGGTTTTCCCAGTCACGAC"
     primer          5622..5643
                     /label="Primer073_MCS-SPEI-pSEVA-for"
                     /note="sequence: CCCTGGCGACTAGTCTTGGACT"
     primer          complement(5627..5649)
                     /label="Primer070_pSEVA-forAbR-USER-rev"
                     /note="sequence: AACAGGAGTCCAAGACTAGTCGC"
     terminator      5636..5738
                     /label="T0"
     primer          5641..5669
                     /label="Primer071_AbR-USER-for"
                     /note="sequence: ACTCCTGTTGATAGATCCAGTAATGACCT"
     primer          complement(5689..5709)
                     /label="Primer063_T0-terminator-seq-rev"
                     /note="sequence: CGGCAACCGAGCGTTCTGAAC"
     primer          complement(5694..5710)
                     /label="Primer109_pS2"
                     /note="sequence: GCGGCAACCGAGCGTTC"
     primer          complement(5705..5734)
                     /label="Primer489_Prom_Term_U_rev"
                     /note="sequence: ATTCTCACCAATAAAAAACGCCCGGCGGCA"
     primer          complement(5711..5738)
                     /label="Primer516_metH_4044-3316_rev"
                     /note="sequence: CTGGATTCTCACCAATAAAAAACGCCCG"
     misc_feature    5746..525
                     /label="Gadget"
     primer_bind     5747..5766
                     /label="pRS-marker"
     CDS             complement(5805..364)
                     /label="SacB"
     CDS             complement(5805..364)
                     /label="Translation 364-5805"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"
     primer          complement(6488..6504)
                     /label="oEO_075"
                     /note="sequence: caaccatacgctgagag"
ORIGIN
        1 aaatcgatgt gtcatccgca tttttaggat ctccggctaa tgcaaagacg atgtggtagc
       61 cgtgatagtt tgcgacagtg ccgtcagcgt tttgtaatgg ccagctgtcc caaacgtcca
      121 ggccttttgc agaagagata tttttaattg tggacgaatc gaactcagga acttgatatt
      181 tttcattttt ttgctgttca gggatttgca gcatatcatg gcgtgtaata tgggaaatgc
      241 cgtatgtttc cttatatggc ttttggttcg tttctttcgc aaacgcttga gttgcgcctc
      301 ctgccagcag tgcggtagta aaggttaata ctgttgcttg ttttgcaaac tttttgatgt
      361 tcatcgttca tgtctccttt tttatgtact gtgttagcgg tctgcttctt ccagccctcc
      421 tgtttgaaga tggcaagtta gttacgcaca ataaaaaaag acctaaaata tgtaaggggt
      481 gacgccaaag tatacacttt gccctttaca cattttaggt cttgcattta aattggcgaa
      541 aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
      601 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
      661 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
      721 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
      781 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
      841 ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg
      901 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
      961 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     1021 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     1081 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     1141 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     1201 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1261 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taatttgact
     1321 tcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     1381 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     1441 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     1501 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     1561 cctgctctgc gaggctggcc gtaggccggc cgcgatgcag gtggctgctg aacccccagc
     1621 cggaactgac cccacaaggc cctagcgttt gcaatgcacc aggtcaccat tgacccaggc
     1681 gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca
     1741 cttcttcacg cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg
     1801 gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt
     1861 gcggtacttc tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc
     1921 ctcgtcgatc aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg
     1981 gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt
     2041 cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca
     2101 gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg
     2161 cttcgcgtac tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac
     2221 gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc
     2281 ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg
     2341 catcgctcgc atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt
     2401 gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc
     2461 caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt
     2521 catcgacttc gccaaacctg ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc
     2581 cgatggcgcg ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt
     2641 agcttgctgg actatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt
     2701 gcggcttgcg atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct
     2761 gtatgccttc cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccggaatt
     2821 aattccccgg atcgatccgt cgatcttgat cccctgcgcc atcagatcct tggcggcaag
     2881 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     2941 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     3001 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     3061 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt ggctgccatt
     3121 tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg
     3181 ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc
     3241 gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta
     3301 aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct
     3361 gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc
     3421 ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata
     3481 ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca
     3541 ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg
     3601 cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc
     3661 tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc ggccctacat
     3721 ggctctgctg tagtgagtgg gttgcgcttc ggcagcggtc ctgatccccc gcagaaaaaa
     3781 aggatctcaa gaagatcctt tgatcttttc tacggcgcgc ccagctgtct agggcggcgg
     3841 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     3901 tttcgactga gcctttcgtt ttatttgatg cctttaatta atgtgacaat taatcatcgg
     3961 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccct aggggagtca
     4021 tgaacatatg agagccctgg tagtcatccg cctgtcccgc gtcaccgatg ctacgacttc
     4081 accggagcgt cagctggagt cttgccagca gctctgcgcc cagcgcggct gggacgtcgt
     4141 cggggtagcg gaggatctgg acgtctccgg ggcggtcgat ccgttcgacc ggaagcgcag
     4201 accgaacctg gcccggtggc tagcgttcga ggagcaaccg tttgacgtga tcgtggcgta
     4261 ccgggtagat cggttgaccc gatcgatccg gcatcttcag cagctggtcc actgggccga
     4321 ggaccacaag aagctggtcg tctccgcgac cgaagcgcac ttcgatacga cgacgccgtt
     4381 tgcggcggtc gtcatcgcgc ttatgggaac ggtggcgcag atggaattag aagcgatcaa
     4441 agagcggaac cgttcggctg cgcatttcaa tatccgcgcc gggaaatacc gaggatccct
     4501 gccgccgtgg ggatacctgc ctacgcgcgt ggacggggag tggcggctgg tgccggaccc
     4561 tgtgcagcga gagcgcatcc tcgaggtgta tcaccgcgtc gtcgacaacc acgagccgct
     4621 gcatctggtg gcccacgacc tgaaccggcg tggtgtcctg tcgccgaagg actacttcgc
     4681 gcagctgcaa ggccgcgagc cgcagggccg ggagtggtcg gctaccgcgc tgaagcgatc
     4741 gatgatctcc gaggcgatgc tcgggtacgc gactctgaac ggtaagaccg tccgagacga
     4801 cgacggagcc ccgctggtgc gggctgagcc gatcctgacc cgtgagcagc tggaggcgct
     4861 gcgcgccgag ctcgtgaaga cctcccgggc gaagcccgcg gtgtctaccc cgtcgctgct
     4921 gctgcgggtg ttgttctgcg cggtgtgcgg ggagcccgcg tacaagttcg ccgggggagg
     4981 acgtaagcac ccgcgctacc gctgccgctc gatggggttc ccgaagcact gcgggaacgg
     5041 cacggtggcg atggccgagt gggacgcgtt ctgcgaggag caggtactgg atctgctcgg
     5101 ggacgcggag cgtctggaga aagtctgggt agcgggctcg gactccgcgg tcgaactcgc
     5161 ggaggtgaac gcggagctgg tggacctgac gtcgctgatc ggctccccgg cctaccgggc
     5221 gggctctccg cagcgagaag cactggatgc ccgtattgcg gcgctggccg cgcggcaaga
     5281 ggagctggag ggcctggagg ctcgcccgtc tggctgggag tggcgcgaga ccgggcagcg
     5341 gttcggggac tggtggcggg agcaggacac cgcggcaaag aacacctggc ttcggtcgat
     5401 gaacgttcgg ctgacgttcg acgtccgcgg cgggctgact cgcacgatcg acttcgggga
     5461 tcttcaggag tacgagcagc atctcaggct cggcagcgtg gtcgaacggc tacacaccgg
     5521 gatgtcgtaa taacgctgat agtgctagtg tagatcgcta cactagtcct agagtcgacc
     5581 tgcaggcatg caagcttgcg gccgcgtcgt gactgggaaa accctggcga ctagtcttgg
     5641 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     5701 cggttgccgc cgggcgtttt ttattggtga gaatccaggg gtcccccggc atcagagcag
     5761 attgtactga gagtgcacca taatcggcat tttcttttgc gtttttattt gttaactgtt
     5821 aattgtcctt gttcaaggat gctgtctttg acaacagatg ttttcttgcc tttgatgttc
     5881 agcaggaagc taggcgcaaa cgttgattgt ttgtctgcgt agaatcctct gtttgtcata
     5941 tagcttgtaa tcacgacatt gtttcctttc gcttgaggta cagcgaagtg tgagtaagta
     6001 aaggttacat cgttaggatc aagatccatt tttaacacaa ggccagtttt gttcagcggc
     6061 ttgtatgggc cagttaaaga attagaaaca taaccaagca tgtaaatatc gttagacgta
     6121 atgccgtcaa tcgtcatttt tgatccgcgg gagtcagtga acagatacca tttgccgttc
     6181 attttaaaga cgttcgcgcg ttcaatttca tctgttactg tgttagatgc aatcagcggt
     6241 ttcatcactt ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt
     6301 gctaactcag ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag
     6361 aatgatgtgc ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca
     6421 tcttcagttc cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag
     6481 tgaggatctc tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc
     6541 tgtacatttt gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg
     6601 ttgatgttca aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt
     6661 ttgccgtaat gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta
     6721 aatgtggctg aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg
     6781 aatttgtcgc tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg
     6841 ccgacttttt gatagaacat gt
//