LOCUS       pCHC004_(∆phaCAB        4491 bp DNA     circular SYN 01-MAY-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4491)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 22, 2024 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          632..1371
                     /chromosome="1"
                     /strain="H16"
                     /mol_type="genomic DNA"
                     /note="Ralstonia H16 chrm 1 AM260479 (1,555,853 - 
                     1,562,703).dna"
                     /db_xref="taxon:381666"
                     /organism="Ralstonia eutropha H16"
     source          1380..2129
                     /chromosome="1"
                     /strain="H16"
                     /mol_type="genomic DNA"
                     /note="Ralstonia H16 chrm 1 AM260479 (1,555,853 - 
                     1,562,703).dna"
                     /db_xref="taxon:381666"
                     /organism="Ralstonia eutropha H16"
     rep_origin      1..595
                     /label=pMB1 replication origin
                     /note="pMB1 replication origin"
     primer_bind     14..31
                     /label=oCJ392
     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..649
                     /label=oCHC021: phaACB Up F + EcoRI Link
                     /note="Assembly of phaACB Upstream Region in pCHC004"
     misc_feature    632..1371
                     /label=phaCAB Upstream Region
                     /note="For phaCAB deletion targeting."
     CDS             632..707
                     /gene="h16_A1435"
                     /locus_tag="H16_A1435"
                     /product="uncharacterized conserved protein"
                     /function="Function unknown"
                     /label=h16_A1435
                     /db_xref="GI:113526225"
                     /db_xref="InterPro:IPR003730"
                     /db_xref="InterPro:IPR011324"
                     /db_xref="UniProtKB/TrEMBL:Q0KBQ1"
                     /inference="protein motif:same species:COG1496"
                     /inference="protein motif:same species:PF02578"
                     /protein_id="CAJ92570.1"
     gene            632..707
                     /gene="h16_A1435"
                     /locus_tag="H16_A1435"
                     /label=h16_A1435
     CDS             complement(721..924)
                     /gene="h16_A1436"
                     /locus_tag="H16_A1436"
                     /product="conserved hypothetical protein"
                     /label=h16_A1436
                     /db_xref="GI:113526226"
                     /db_xref="UniProtKB/TrEMBL:Q0KBQ0"
                     /protein_id="CAJ92571.1"
     gene            complement(721..924)
                     /gene="h16_A1436"
                     /locus_tag="H16_A1436"
                     /label=h16_A1436
     primer_bind     complement(1348..1401)
                     /label=oCHC022: phaACB Up R + Pme1 Link
                     /note="Assembly of phaACB Upstream Region in pCHC004"
     primer_bind     1350..1394
                     /label=oCHC023: phaACB Down F + Pme1 Link
                     /note="Assembly  of phaACB Downstream Region in pCHC004"
     misc_feature    1372..1379
                     /label=Link
                     /note="Introducing Pme1 site for creating knock-in 
                     vectors."
     misc_feature    1380..2129
                     /label=phaCAB Downstream Region
                     /note="For phaCAB deletion targeting."
     CDS             1816..2129
                     /gene="phaR"
                     /locus_tag="H16_A1440"
                     /product="transcriptional regulator of phasin expression"
                     /function="Transcription"
                     /label=phaR
                     /db_xref="GI:113526230"
                     /db_xref="InterPro:IPR007897"
                     /db_xref="InterPro:IPR010134"
                     /db_xref="InterPro:IPR012909"
                     /db_xref="UniProtKB/TrEMBL:Q0KBP6"
                     /inference="protein motif:same species:COG5394"
                     /protein_id="CAJ92575.1"
     gene            1816..2129
                     /gene="phaR"
                     /locus_tag="H16_A1440"
                     /label=phaR
     primer_bind     complement(2109..2159)
                     /label=oCHC024: phaACB Down R + HindIII Link
                     /note="Assembly  of phaACB Downstream Region in pCHC004"
     primer_bind     complement(2139..2156)
                     /label=M13F
                     /note="M13F"
     promoter        2166..2215
                     /label=NEOKAN Promoter
     -35_signal      2172..2177
                     /label=-35
     primer_bind     complement(2189..2207)
                     /label=oCJ681
     -10_signal      2195..2200
                     /label=-10
     primer_bind     complement(2225..2250)
                     /label=oCHC026: pK18sB seq R
                     /note="Sequencing  inserts in pK18sB MCS."
     primer_bind     complement(2234..2253)
                     /label=oCJ547
     CDS             2237..3031
                     /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"
     gene            2237..3031
                     /gene="nptII"
                     /label=nptII
     primer_bind     2546..2564
                     /label=oCJ389
     gene            3038..4459
                     /gene="sacB"
                     /label=sacB
     CDS             3038..4459
                     /codon_start=1
                     /gene="sacB"
                     /product="Llevan 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"
     primer_bind     3191..3209
                     /label=oCJ390
     primer_bind     3829..3847
                     /label=oCJ391
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 ccgccggtcg cttctactcc tatcggcgcg
      661 atggcgtgac cggccgcatg gccagcctgg tctggctggc ggactgagcc cgccgctgcc
      721 tcactcgtcc ttgcccctgg ccgcctgcgc gcgctcggct tcagccttgc gtcggcggcg
      781 gccgggcgtg cccatgatgt agagcaccag cgccaccggc gccatgccat acatcaggaa
      841 ggtggcaacg cctgccacca cgttgtgctc ggtgatcgcc atcatcagcg ccacgtagag
      901 ccagccaatg gccacgatgt acatcaaaaa ttcatccttc tcgcctatgc tctggggcct
      961 cggcagatgc gagcgctgca taccgtccgg taggtcggga agcgtgcagt gccgaggcgg
     1021 attcccgcat tgacagcgcg tgcgttgcaa ggcaacaatg gactcaaatg tctcggaatc
     1081 gctgacgatt cccaggtttc tccggcaagc atagcgcatg gcgtctccat gcgagaatgt
     1141 cgcgcttgcc ggataaaagg ggagccgcta tcggaatgga cgcaagccac ggccgcagca
     1201 ggtgcggtcg agggcttcca gccagttcca gggcagatgt gccggcagac cctcccgctt
     1261 tgggggaggc gcaagccggg tccattcgga tagcatctcc ccatgcaaag tgccggccag
     1321 ggcaatgccc ggagccggtt cgaatagtga cggcagagag acaatcaaat cgtttaaacc
     1381 ctgccggcct ggttcaacca gtcggcagcc ggcgctggcg cccgcgtatt gcggtgcagc
     1441 cagcgcggcg cacaaggcgg cgggcgtttc gtttcgccgc ccgtttcgcg ggccgtcaag
     1501 gcccgcgaat cgtttctgcc cgcgcggcat tcctcgcttt ttgcgccaat tcaccgggtt
     1561 ttccttaagc cccgtcgctt ttcttagtgc cttgttgggc atagaatcag ggcagcggcg
     1621 cagccagcac catgttcgtg cagcgcggcc ctcgcggggg cgaggctgca ggccgccacg
     1681 cgcagccatg cgcgaacggg ccaccagatg gccggcacga caacaagcag atggcgcggg
     1741 cgataccgat ttgcgcactg caccccatgc ggtgcagcag cgcgcaaaca gcgatgacac
     1801 aaggacagag caccgatggc cacgaccaaa aaaggcgcag agcgactgat caaaaagtat
     1861 ccgaaccgta ggctctacga cacccagacc agcacctaca tcaccctggc cgacgtcaag
     1921 cagctggtca tggattcaga agaattcaag gtcgtcgacg ccaagtctgg tgacgaactg
     1981 acccgcagca tcttgctgca gatcatcctg gaagaagaaa cgggcggcgt gccgatgttc
     2041 tccagcgcga tgctgtcgca gatcatccgc ttctacggcc atgccatgca gggcatgatg
     2101 ggcacctacc tggaaaagaa catccaggca agcttggcac tggccgtcgt tttacaacgt
     2161 cgtgaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaacagg
     2221 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2281 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2341 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2401 tgccctgaat gaactccaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2461 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2521 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2581 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2641 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2701 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2761 ggcgcggatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     2821 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2881 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     2941 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3001 cttctatcgc cttcttgacg agttcttctg agcgacgatg aacatcaaaa agtttgcaaa
     3061 acaagcaaca gtattaacct ttactaccgc actgctggca ggaggcgcaa ctcaagcgtt
     3121 tgcgaaagaa acgaaccaaa agccatataa ggaaacatac ggcatttccc atattacacg
     3181 ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag tttctgaatt
     3241 tgattcgtcc acaattaaaa atatctcttc tgcaaaaggc ctggacgttt gggacagctg
     3301 gccattacaa aacgctgacg gcactgtcgc aaactatcac ggctaccaca tcgtctttgc
     3361 attagccgga gatcctaaaa atgcggatga cacatcgatt tacatgttct atcaaaaagt
     3421 cggcgaaact tctattgaca gctggaaaaa cgctggccgc gtctttaaag acagcgacaa
     3481 attcgatgca aatgattcta tcctaaaaga ccaaacacaa gaatggtcag gttcagccac
     3541 atttacatct gacggaaaaa tccgtttatt ctacactgat ttctccggta aacattacgg
     3601 caaacaaaca ctgacaactg cacaagttaa cgtatcagca tcagacagct ctttgaacat
     3661 caacggtgta gaggattata aatcaatctt tgacggtgac ggaaaaacgt atcaaaatgt
     3721 acagcagttc atcgatgaag gcaactacag ctcaggcgac aaccatacgc tgagagatcc
     3781 tcactacgta gaagataaag gccacaaata cttagtattt gaagcaaaca ctggaactga
     3841 agatggctac caaggcgaag aatctttatt taacaaagca tactatggca aaagcacatc
     3901 attcttccgt caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca cggctgagtt
     3961 agcaaacggc gctctcggta tgattgagct aaacgatgat tacacactga aaaaagtgat
     4021 gaaaccgctg attgcatcta acacagtaac agatgaaatt gaacgcgcga acgtctttaa
     4081 aatgaacggc aaatggtacc tgttcactga ctcccgcgga tcaaaaatga cgattgacgg
     4141 cattacgtct aacgatattt acatgcttgg ttatgtttct aattctttaa ctggcccata
     4201 caagccgctg aacaaaactg gccttgtgtt aaaaatggat cttgatccta acgatgtaac
     4261 ctttacttac tcacacttcg ctgtacctca agcgaaagga aacaatgtcg tgattacaag
     4321 ctatatgaca aacagaggat tctacgcaga caaacaatca acgtttgcgc cgagcttcct
     4381 gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca
     4441 attaacagtt aacaaataat cagaccccgt agaaaagatc aaaggatctt c
//
