LOCUS       sequence-484959-        2952 bp    DNA     circular SYN 28-AUG-2025
DEFINITION  Expresses influenza A virus mvRNA PA-66.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2952)
  TITLE     mvRNA expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2952)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pPolI-PA-66
FEATURES             Location/Qualifiers
     source          1..2952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..88
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(103..108,109..126,127..133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..157
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     146..168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     165..181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     165..181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(657..674)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(657..673)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(666..688)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(882..901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1001..1023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1061..1079)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1147..1251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1252..1320,1321..2112)
                     /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(1470..1489)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2283..2871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2772..2791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
       61 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      121 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      181 catgattacg aattcgagct cggtacccgg ggatcctcta gagtcccatt cgccattacc
      241 gaggggacgg tcccctcgga atgttgccca gccggcgcca gcgaggaggc tgggaccatg
      301 ccggccagcg aaagcaggta ctgattcaaa atgtgctatc catactgtcc aaaaaagtac
      361 cttgtttcta ctaataaccc ggcggcccaa aatgccgact cggagcgaaa gatatacctc
      421 ccccggggcc gggaggtcgc gtcaccgacc acgccgccgg cccaggcgac gcgcgacacg
      481 gacacctgtc cccaaaaacg ccaccatcgc agccacacac ggagcgcccg gggccctctg
      541 gtcaacccca ggacacacgc gggagcagcg ccgggccggg gacgccctcc cggccgcccg
      601 tgccacacgc aggggccggc ccgtctagaa ttgacctgca ggcatgcaag cttggcactg
      661 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      721 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      781 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
      841 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
      901 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1021 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     1081 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1141 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1201 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1261 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1321 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1381 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1441 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1501 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1561 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1621 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1681 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1741 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     1801 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1861 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1921 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     1981 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2041 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2101 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2161 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2221 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2281 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2341 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2401 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2461 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2521 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2581 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2641 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2701 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2761 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2821 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2881 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2941 gcgttatccc ct
//
