LOCUS       sequence-375584-        8950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The PhiC31-phage based integration vector containing the actII-ORF4 
            gene under the control of strong kasOp* promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8950)
  AUTHORS   Novakova R, Homerova D, Csolleiova D, Rezuchova B, Sevcikova B, 
            Javorova R, Feckova L, Kormanec J
  TITLE     A stable vector for efficient production of heterologous proteins 
            and secondary metabolites in streptomycetes.
  JOURNAL   Appl Microbiol Biotechnol. 2022 Nov;106(21):7285-7299. doi: 
            10.1007/s00253-022-12187-4. Epub 2022 Sep 29.
  PUBMED    36173451
REFERENCE   2  (bases 1 to 8950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1007/s00253-022-12187-4"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2022-11"; volume: "106"; issue: "21"; pages: 
            "7285-7299"
COMMENT     Sequence Label: pPhiC31-actIIORF4
FEATURES             Location/Qualifiers
     source          1..8950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            252..361
                     /note="incP origin of transfer"
     CDS             871..1665
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(925..944)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1535..1554
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    1692..1791
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             1801..3618
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3779..3847,3848..4639)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3997..4016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5552..5569
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5584..5724
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5650..5669)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5810..5832
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5826..6017)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDTDEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7475..7494
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gggtcaggcc gcgggggttc ctccggccgc ccgctcggcc tccggcgcgg cttcctgcgc
       61 ggccgcctcg gcggcgcggc gctccccctc cgcctgggaa cgcttgccgt agcgggccgt
      121 ccacagcgcc aggccgccca gcgtcccgta gatcatgaac gggctgatcg aattcgttaa
      181 cgtataccaa ttgctggttg gcttggtttc atcagccatc cgcttgccct catctgttac
      241 gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga
      301 ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc
      361 cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt
      421 atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg
      481 ttatgcagcg gaaaagatcc gtcgacctgc aggatagctt cacgctgccg caagcactca
      541 gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc
      601 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
      661 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg
      721 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
      781 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
      841 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
      901 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
      961 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     1021 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     1081 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     1141 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     1201 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     1261 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     1321 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     1381 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     1441 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     1501 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     1561 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1621 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctcttaa
     1681 gttcacgttt tcccaggtca gaagcggttt tcgggagtag tgccccaact ggggtaacct
     1741 ttgagttctc tcagttgggg gcgtagggtc gccgacatga cacaaggggt tgtgaccggg
     1801 gtggacacgt acgcgggtgc ttacgaccgt cagtcgcgcg agcgcgaaaa ttcgagcgca
     1861 gcaagcccag cgacacagcg tagcgccaac gaagacaagg cggccgacct tcagcgcgaa
     1921 gtcgagcgcg acgggggccg gttcaggttc gtcgggcatt tcagcgaagc gccgggcacg
     1981 tcggcgttcg ggacggcgga gcgcccggag ttcgaacgca tcctgaacga atgccgcgcc
     2041 gggcggctca acatgatcat tgtctatgac gtgtcgcgct tctcgcgcct gaaggtcatg
     2101 gacgcgattc cgattgtctc ggaattgctc gccctgggcg tgacgattgt ttccactcag
     2161 gaaggcgtct tccggcaggg aaacgtcatg gacctgattc acctgattat gcggctcgac
     2221 gcgtcgcaca aagaatcttc gctgaagtcg gcgaagattc tcgacacgaa gaaccttcag
     2281 cgcgaattgg gcgggtacgt cggcgggaag gcgccttacg gcttcgagct tgtttcggag
     2341 acgaaggaga tcacgcgcaa cggccgaatg gtcaatgtcg tcatcaacaa gcttgcgcac
     2401 tcgaccactc cccttaccgg acccttcgag ttcgagcccg acgtaatccg gtggtggtgg
     2461 cgtgagatca agacgcacaa acaccttccc ttcaagccgg gcagtcaagc cgccattcac
     2521 ccgggcagca tcacggggct ttgtaagcgc atggacgctg acgccgtgcc gacccggggc
     2581 gagacgattg ggaagaagac cgcttcaagc gcctgggacc cggcaaccgt tatgcgaatc
     2641 cttcgggacc cgcgtattgc gggcttcgcc gctgaggtga tctacaagaa gaagccggac
     2701 ggcacgccga ccacgaagat tgagggttac cgcattcagc gcgacccgat cacgctccgg
     2761 ccggtcgagc ttgattgcgg accgatcatc gagcccgctg agtggtatga gcttcaggcg
     2821 tggttggacg gcagggggcg cggcaagggg ctttcccggg ggcaagccat tctgtccgcc
     2881 atggacaagc tgtactgcga gtgtggcgcc gtcatgactt cgaagcgcgg ggaagaatcg
     2941 atcaaggact cttaccgctg ccgtcgccgg aaggtggtcg acccgtccgc acctgggcag
     3001 cacgaaggca cgtgcaacgt cagcatggcg gcactcgaca agttcgttgc ggaacgcatc
     3061 ttcaacaaga tcaggcacgc cgaaggcgac gaagagacgt tggcgcttct gtgggaagcc
     3121 gcccgacgct tcggcaagct cactgaggcg cctgagaaga gcggcgaacg ggcgaacctt
     3181 gttgcggagc gcgccgacgc cctgaacgcc cttgaagagc tgtacgaaga ccgcgcggca
     3241 ggcgcgtacg acggacccgt tggcaggaag cacttccgga agcaacaggc agcgctgacg
     3301 ctccggcagc aaggggcgga agagcggctt gccgaacttg aagccgccga agccccgaag
     3361 cttccccttg accaatggtt ccccgaagac gccgacgctg acccgaccgg ccctaagtcg
     3421 tggtgggggc gcgcgtcagt agacgacaag cgcgtgttcg tcgggctctt cgtagacaag
     3481 atcgttgtca cgaagtcgac tacgggcagg gggcagggaa cgcccatcga gaagcgcgct
     3541 tcgatcacgt gggcgaagcc gccgaccgac gacgacgaag acgacgccca ggacggcacg
     3601 gaagacgtag cggcgtagcg agacacccgc ggccgccacc gcggtggcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4021 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4321 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5461 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5521 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5581 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5641 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     5701 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     5761 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5821 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     5881 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgtgtc cagctcgttg
     5941 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     6001 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     6061 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     6121 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     6181 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     6241 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     6301 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     6361 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     6421 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     6481 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     6541 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     6601 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     6661 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     6721 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     6781 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     6841 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     6901 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     6961 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     7021 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     7081 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     7141 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     7201 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     7261 ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     7321 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     7381 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     7441 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     7501 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctaca
     7561 ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca
     7621 ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt
     7681 gcatcaacgc atatagcgct aggcgcgcca cagctgttcc ggggatccgt cgacccggta
     7741 ctctagactc gaggatatcg gcatcgatgc atggatccat tcgaacggtc tctgctttga
     7801 caacatgctg tgcggtgttg taaagtcgtg gccaggagat gactgactga ggtaccagat
     7861 ctgactgagt gaccaaagga ggcggacata tgagattcaa cttattggga cgtgtccatg
     7921 taatcaccga tgcgggatgt gtaattccgc ttaaatcctc gaaggcgacc cagctcctgg
     7981 tgctgctgct cctcaggcgg cacgaggtgg tgggatcggg ggtgctcatc gaggagttgt
     8041 gggcggacca cccgccccgc agcgccatga cgacgctgca gacgtacgtg taccacaccc
     8101 gccggctgct gggggagcac cgggtgacga gcgacgaccg ggaattggtc ctgacccagc
     8161 cgcccggcta cttcgccctg atcgacgagg acgaactcga cgtcgcggtc gccgagcgtc
     8221 tgatccgcac cggcggccgg ctgctcgagg agaaccggct cgaggaggcg ctcgcctcgt
     8281 tggacgcggg actggatctc tggcgaggcc cggcgctgtc caccgtaccg tgcggccggg
     8341 tgctcgaaag caatatcgcg cacctggaag agctgcggct ttttggaatg cagctccgta
     8401 tcgacgcgaa ttggcggctg ggcagaatag ggccgatgat tccggaactc cggtccctgg
     8461 taatttcgca tccgctgaac gagaccctgc acgccaaact gatgggcgcg ctctgtcaga
     8521 tgggcaggcg cgccgaggcg ctggaatcgt atcggaatct ccggcggata ctgtccgacg
     8581 aactgggggt ggatccgacg ccggaaatcc agcgtatgca catggaaatt ctcaacggtg
     8641 agaaggtgct cgtgtagcaa gcttgttctg gaacatcgag ttcgacgacg ccggcgccat
     8701 cggccgccgc taccgccgcc aggacgagat cggtacgccg ttctgcgtca ccgtcgactt
     8761 cgacaccctg gacgacaacg cggtgaccgt gcgcgagcgc gacaccatga agcaggagcg
     8821 cgtctccctg gaccagatcc agagctacct cggcagccgt ctgctcggct gctaggtctc
     8881 cacgcgaaaa ggccggtggc ccggtgcgcg atccgcgcac cgggccaccg gctttcgtgt
     8941 gcttcgcggc
//
