LOCUS       pCHC027_(∆pemK)        4737 bp DNA     circular SYN 15-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4737)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 22, 2024 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..595
                     /label=pMB1 replication origin
                     /note="pMB1 replication origin"
     primer_bind     14..31
                     /label=oCJ392
     primer_bind     complement(110..138)
                     /label=oCHC106:  OriT Seq R
                     /note="Sequencing of RP4 OriT."
     primer_bind     510..531
                     /label=oCHC025: pK18sB seq F
                     /note="Sequencing  inserts in pK18sB MCS."
     primer_bind     510..527
                     /label=oCJ546
     primer_bind     553..571
                     /label=oCJ680
     primer_bind     601..617
                     /label=M13R
     primer_bind     602..658
                     /label=oCHC154:  pemK Up F + EcoRI
                     /note="Assembly of pemK ko plasmid."
     misc_feature    632..1381
                     /label=pemK UP
     gene            1127..1381
                     /gene="pemI"
                     /locus_tag="PHG380"
                     /label=pemI
     CDS             1127..1381
                     /codon_start=1
                     /transl_table=11
                     /gene="pemI"
                     /locus_tag="PHG380"
                     /product="putative PemI-like protein
                     antitoxin MazE "
                     /function="plasmid maintenance postsegregational 
                     killingmechanism"
                     /label=PemI
                     /label=putative PemI-like protein
                     /note="similar to Nostoc sp. pcc 7120 gi:17230704"
                     /db_xref="GI:32527379"
                     /protein_id="AAP86129.1"
                     /translation="MRKSATLTIQKWGNSLAVRIPTAVARSAHFAEGQEVEVSVDEIGV
                     TVRPVGRRALTLAEKLALFDPIKHGGEAMATQRVGAEAM"
     primer_bind     1352..1405
                     /label=oCHC156:  pemK Dn F + Link
                     /note="Assembly of pemK ko plasmid."
     primer_bind     complement(1358..1411)
                     /label=oCHC155:  pemK Up R + Link
                     /note="Assembly of pemK ko plasmid."
     gene            1381
                     /gene="pemK"
                     /locus_tag="PHG381"
                     /label=pemK
     CDS             1381
                     /codon_start=1
                     /transl_table=11
                     /gene="pemK"
                     /locus_tag="PHG381"
                     /product="putative PemK-like protein
                     mRNA interferase MazF "
                     /function="plasmid maintenance, postsegregational 
                     killingsystem"
                     /label=PemK
                     /label=putative PemK-like protein
                     /note="similar to Deinococcus radiodurans gi:15805444"
                     /db_xref="GI:32527380"
                     /protein_id="AAP86130.1"
                     /translation=""
     misc_feature    1382..2131
                     /label=pemK DN
     gene            complement(1382..1743)
                     /gene="nagM6"
                     /locus_tag="PHG382"
                     /label=nagM6
     CDS             complement(1382..1743)
                     /codon_start=1
                     /transl_table=11
                     /gene="nagM6"
                     /locus_tag="PHG382"
                     /product="putative NagM-like protein"
                     /label=putative NagM-like protein
                     /note="Pfam entry UPF0065; similar to 
                     Agrobacteriumtumefaciens gi:15890266"
                     /db_xref="GI:32527381"
                     /protein_id="AAP86131.1"
                     /translation="MTSARGPRGATSPAHCPALATQAKAQLQKITYATPGIGTLLHLIG
                     VVLDKNSSVPLQHVPYKGAGSAINDLLGGQVDVLITSTSTVAGFIQSSRMCALAVTSLR
                     RLGVFAKTTRHAVKHG"
     primer_bind     complement(2092..2151)
                     /label=oCHC157:  pemK Dn R + HindIII
                     /note="Assembly of pemK ko plasmid."
     gene            complement(2121..2131)
                     /gene="tutC"
                     /locus_tag="PHG383"
                     /label=tutC
     primer_bind     complement(2141..2158)
                     /label=M13F
                     /note="M13F"
     promoter        2168..2217
                     /label=NEOKAN Promoter
     primer_bind     complement(2191..2209)
                     /label=oCJ681
     primer_bind     complement(2227..2252)
                     /label=oCHC026: pK18sB seq R
                     /note="Sequencing  inserts in pK18sB MCS."
     primer_bind     complement(2236..2255)
                     /label=oCJ547
     gene            2239..3033
                     /gene="nptII"
                     /label=nptII
     CDS             2239..3033
                     /codon_start=1
                     /gene="nptII"
                     /product="Neomycin Phosphotransferase"
                     /label=KanR (neomycin phosphotransferase)
                     /label=neomycin phosphotransferase
                     /db_xref="GI:215434895"
                     /protein_id="ACJ66844.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     2548..2566
                     /label=oCJ389
     CDS             3040..4461
                     /codon_start=1
                     /gene="sacB"
                     /product="Levan Sucrase"
                     /label=SacB (levan sucrase)
                     /label=levan sucrase
                     /db_xref="GI:215434896"
                     /protein_id="ACJ66845.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            3040..4461
                     /gene="sacB"
                     /label=sacB
     primer_bind     3193..3211
                     /label=oCJ390
     primer_bind     3831..3849
                     /label=oCJ391
     primer_bind     4309..4352
                     /label=oCHC105:  OriT Seq F
                     /note="Sequencing of RP4 OriT."
     primer_bind     complement(4435..4477)
                     /label=oCHC108: pK18sB R
                     /note="For linearization of pK18sB."
     primer_bind     complement(4439..4599)
                     /label=oCHC104:  pk18sB+OriT R
                     /note="For Q5 insertion of OriT into pk18sB."
     primer_bind     4453..4497
                     /label=oCHC109:  OriT F + pK18sB Link
                     /note="For amplification/insertion of OriT into pK18sB."
     rep_origin      4478..4721
                     /label=RP4 OriT
                     /note="RP4 OriT"
     primer_bind     join(4600..4737,1..20)
                     /label=oCHC103: pk18sB+OriT F
                     /note="For Q5 insertion of OriT into pk18sB."
     primer_bind     complement(join(4695..4737,1..9))
                     /label=oCHC110:  OriT R + pK18sB Link
                     /note="For amplification/insertion of OriT into pK18sB."
     primer_bind     join(4722..4737,1..22)
                     /label=oCHC107: pK18sB F
                     /note="For linearization of pK18sB."
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgcctcaca
      601 caggaaacag ctatgacatg attacgaatt cagctgctac ctcgaggctg cacaagagat
      661 tcgagccgat tgccataacc cactgcgaca ggtcacctgg taggggctca gcgtcgcgcc
      721 aagtgtggca gccgtggcgt cggccagcga agaaagcgcc ccgcggcggt gcagatttcc
      781 ggcgggttgc cgagaaagga ggctcaaatg tcctcacagg aaaacagcgg ccatgtcgaa
      841 caaggcaatg gcaaccgtgt cgaagctggc gctagcgggg ctgcggcatg gcgccggtgc
      901 ggtagccgga gcccggcgcg gcgccaagca gatacgcagg cttctgccgg cagactcctt
      961 cgcgcgttcg gctgcgcagc gccgctgcgc cgtcagtcgt cacgcgggca aatttcattt
     1021 gttggcagcg atggcgagca acgctcgagg gagcgatgta gatacgcgtt gaaacatgga
     1081 tctcttatgt ttatacttgt atcaacattg tttggaggca tctattatgc gaaaaagcgc
     1141 aaccctgacg attcaaaagt ggggcaacag cttggcggtt cgaatcccca ctgcggtggc
     1201 tcgttctgca catttcgccg agggccagga agtggaggta tccgtcgatg agattggcgt
     1261 aactgttcga ccagttggtc gtcgtgccct cactctcgcg gaaaagcttg ctctgtttga
     1321 ccccatcaag cacggcggcg aagctatggc cacgcagcgt gtcggcgcgg aggccatgta
     1381 acctccatgc ttgaccgcat ggcgcgttgt cttggcgaac acgccgagcc gtcgcaagct
     1441 ggtcactgcc agcgcacaca tgcggctcga ctggatgaag ccagccacgg tagacgtaga
     1501 cgtgatcagc acatcaacct gaccaccgag cagatcattg atggccgacc cggcgccctt
     1561 gtacggcacg tgctgcagcg gcacgctgct gttcttgtcg agcacaacgc ctatcaggtg
     1621 caacagcgtg ccgatgccgg gcgtggcgta ggtgatcttc tgcagttgtg ccttggcctg
     1681 tgtggccagc gccgggcagt gggcgggact cgtcgcgccg cgcggtccgc gtgccgaagt
     1741 cactgcgccg cccaaggttc gcgctgcgga actgggcgcc ggcctgcgtg agcgtcaatg
     1801 ctggcggcgg cccccccggc agcatggcgc gccgccaacc ccatgttgag gttgtgtccc
     1861 ggcgctactg gtcaacgcgc acaatcaacc tcacctggcg cacaacctac ctcaccgatc
     1921 cctgccggct gtttccttcc agcctttgtc cagcttggga gataagacat atgcacaggt
     1981 cacgcacaat acatctcacc ttaggggcat caacacaaca aacctcacct tcttggggcg
     2041 gcttcggatg cggtgccgtt catcaggcat cgtgtccgcc gtaacgggga tgtcgatgta
     2101 caaacagtag tcggcaaaca ctatgcaaat aaagcttggc actggccgtc gttttacaac
     2161 gtcgtgaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaaca
     2221 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2281 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2341 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2401 ggtgccctga atgaactcca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2461 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2521 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2581 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2641 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2701 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2761 aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2821 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2881 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2941 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3001 gccttctatc gccttcttga cgagttcttc tgagcgacga tgaacatcaa aaagtttgca
     3061 aaacaagcaa cagtattaac ctttactacc gcactgctgg caggaggcgc aactcaagcg
     3121 tttgcgaaag aaacgaacca aaagccatat aaggaaacat acggcatttc ccatattaca
     3181 cgccatgata tgctgcaaat ccctgaacag caaaaaaatg aaaaatatca agtttctgaa
     3241 tttgattcgt ccacaattaa aaatatctct tctgcaaaag gcctggacgt ttgggacagc
     3301 tggccattac aaaacgctga cggcactgtc gcaaactatc acggctacca catcgtcttt
     3361 gcattagccg gagatcctaa aaatgcggat gacacatcga tttacatgtt ctatcaaaaa
     3421 gtcggcgaaa cttctattga cagctggaaa aacgctggcc gcgtctttaa agacagcgac
     3481 aaattcgatg caaatgattc tatcctaaaa gaccaaacac aagaatggtc aggttcagcc
     3541 acatttacat ctgacggaaa aatccgttta ttctacactg atttctccgg taaacattac
     3601 ggcaaacaaa cactgacaac tgcacaagtt aacgtatcag catcagacag ctctttgaac
     3661 atcaacggtg tagaggatta taaatcaatc tttgacggtg acggaaaaac gtatcaaaat
     3721 gtacagcagt tcatcgatga aggcaactac agctcaggcg acaaccatac gctgagagat
     3781 cctcactacg tagaagataa aggccacaaa tacttagtat ttgaagcaaa cactggaact
     3841 gaagatggct accaaggcga agaatcttta tttaacaaag catactatgg caaaagcaca
     3901 tcattcttcc gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg cacggctgag
     3961 ttagcaaacg gcgctctcgg tatgattgag ctaaacgatg attacacact gaaaaaagtg
     4021 atgaaaccgc tgattgcatc taacacagta acagatgaaa ttgaacgcgc gaacgtcttt
     4081 aaaatgaacg gcaaatggta cctgttcact gactcccgcg gatcaaaaat gacgattgac
     4141 ggcattacgt ctaacgatat ttacatgctt ggttatgttt ctaattcttt aactggccca
     4201 tacaagccgc tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc taacgatgta
     4261 acctttactt actcacactt cgctgtacct caagcgaaag gaaacaatgt cgtgattaca
     4321 agctatatga caaacagagg attctacgca gacaaacaat caacgtttgc gccgagcttc
     4381 ctgctgaaca tcaaaggcaa gaaaacatct gttgtcaaag acagcatcct tgaacaagga
     4441 caattaacag ttaacaaata atcagacccc gtagaaacgg ccagcctcgc agagcaggat
     4501 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg
     4561 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta
     4621 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa
     4681 tcgctataat gaccccgaag cagggttatg cagcggaaaa gagatcaaag gatcttc
//
