LOCUS       sequence-360816-        5177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for production of Phi6 P2 RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5177)
  AUTHORS   Gregorova P, Heinonen MK, Sarin LP
  TITLE     An improved RT-qPCR method for direct quantification of enveloped 
            RNA viruses.
  JOURNAL   MethodsX. 2022 May 23;9:101737. doi: 10.1016/j.mex.2022.101737. 
            eCollection 2022.
  PUBMED    35669085
REFERENCE   2  (bases 1 to 5177)
  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"; journalName: "MethodsX.";
            date: "2022-05-23"; volume: "9:101737. doi"; pages: " 
            10.1016/j.mex.2022.101737. eCollection 2022"
COMMENT     Sequence Label: T7-Phi6-P2
FEATURES             Location/Qualifiers
     source          1..5177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..32
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     113..132
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        113..131
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2816..2835)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3049..3068)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3228..3246)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3314..3418
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3419..3487,3488..4279)
                     /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     complement(3637..3656)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4450..5038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4939..4958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
       61 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcttatcgaa attaatacga
      121 ctcactatag ggagacccaa gcttccaata ctaccggaaa attcgtgtca aaggatcgta
      181 tggtcgctga cttcgaatac gcagtcacgg gcggtgagca aggctcgctg ttcgctgctt
      241 cgaaggatgc ctctcgtttg aaggaacagt acgggataga tgtcccggac gggtttttct
      301 gcgagcggcg tcgtaccgct atgggtggtc cgttcgcgtt gaacgctcct atcatggccg
      361 ttgcgcaacc tgtgcgaaac aaaatttact ccaagtacgc ttacaccttt caccatacta
      421 ctcgtcttaa taaggaggaa aaggtgaaag agtggtcgtt gtgcgtcgct actgacgtat
      481 ccgaccacga cacgttctgg cctggatggc tgcgggatct catctgtgat gaactgctca
      541 acatggggta cgctccgtgg tgggttaagt tgttcgagac ctcgctcaaa ctgcccgttt
      601 acgtgggcgc tcctgctcct gagcagggcc acacgttgtt gggtgatccg tccaaccctg
      661 atctcgaagt tggtctctcg tccggacaag gggcgaccga cctcatgggc acgttgctca
      721 tgagtatcac ctacctggtg atgcaacttg atcacaccgc tcctcacctc aacagtcgaa
      781 tcaaggacat gccatcagca tgccgctttc ttgactcgta ttggcaagga cacgaggaga
      841 tccgtcagat ctcaaaatct gatgatgcta tgcttggctg gaccaaaggt cgtgctttgg
      901 ttggtggtca tcgtttgttc gagatgctga aagagggtaa ggttaacccc tcaccttaca
      961 tgaagatctc ctacgagcac ggtggcgcct tccttggtga catcctgctt tacgactcgc
     1021 gtcgtgagcc tggctctgcc atcttcgttg gtaacatcaa ctcaatgctg aacaaccagt
     1081 tcagccctga gtacggtgtc caatcgggcg ttcgcgaccg atctaagcgc aaacggccgt
     1141 tccccggtct tgcttgggcg tcgatgaaag atacctacgg tgcctgtccg atctactctg
     1201 atgtgctgga ggcgatcgag cgttgctggt ggaacgcgtt cggtgagtcg taccgtgcgt
     1261 atcgtgaaga tatgcttaaa cgcgacactc tcgaactatc acgctacgtt gcgtcgatgg
     1321 ctcgtcaagc cgggctggct gaactcactc ccattgattt ggaggtgctt gctgacccga
     1381 acaaactcca gtataagtgg accgaggccg atgtctcggc gaatatccac gaggtactga
     1441 tgcatggcgt atcggtcgaa aagactgagc gctttctccg ttctgtaatg ccgaggtaat
     1501 catgccgatt gtcgtaactc aagcgcatat tgatcgtgtc ggcatcgccg ccgatctgct
     1561 cgatgcgtct cctgtgtcgc ttcaagttct tggtcgccct accagtattg gcgaattcct
     1621 ctggatctac tgggttacct aagggtgtgg cccttccgca tagaactgcc tgcgtcagat
     1681 tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa
     1741 gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg
     1801 gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttttacga tcccttcagg
     1861 attacaaaat tcaaagtgcg ttgctagtac caaccctatt ttcattcttc gccaaaagca
     1921 ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggg ggcgcacctc
     1981 tttcgaaaga agtcggggaa gcggttgcaa aacgcttcca tcttccaggg atacgacaag
     2041 gatatgggct cactgagact acatcagcta ttctgattac acccgagggg gatgataaac
     2101 cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg
     2161 ggaaaacgct gggcgttaat cagagaggcg aattatgtgt cagaggacct atgattatgt
     2221 ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac
     2281 attctggaga catagcttac tgggacgaag acgaacactt cttcatagtt gaccgcttga
     2341 agtctttaat taaatacaaa ggatatcagg tggcccccgc tgaattggaa tcgatattgt
     2401 tacaacaccc caacatcttc gacgcgggcg tggcaggtct tcccgacgat gacgccggtg
     2461 aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg
     2521 tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg
     2581 tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc
     2641 tcataaaggc caagaagggc ggaaagtcca aattgtaaaa tgtaactgta ttcagcgatg
     2701 acgaaattct tagctattgt aatcctccga gggggcgagc tcccaaaaaa aaaaaaaaaa
     2761 aaaaaaaaaa aaaaccgaat tggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     2821 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     2881 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
     2941 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
     3001 ctaatgcagg agtcgcataa gggagagcgt cgaatggtgc actctcagta caatctgctc
     3061 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3121 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3181 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3241 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3301 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3361 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3421 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3481 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3541 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3601 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3661 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3721 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3781 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3841 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3901 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3961 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4021 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4081 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4141 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4201 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4261 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4321 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4381 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4441 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4501 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4561 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4621 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4681 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4741 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4801 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4861 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4921 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4981 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5041 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5101 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5161 taccgctcgc cgcagcc
//
