LOCUS       sequence-201044-        3578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2V6 cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3578)
  AUTHORS   Tutucci E, Vera M, Biswas J, Garcia J, Parker R, Singer RH
  TITLE     An improved MS2 system for accurate reporting of the mRNA life 
            cycle.
  JOURNAL   Nat Methods. 2017 Nov 13. doi: 10.1038/nmeth.4502.
  PUBMED    29131164
REFERENCE   2  (bases 1 to 3578)
  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: "Nat 
            Methods. 2017 Nov 13. doi: 10.1038/nmeth.4502."
COMMENT     Sequence Label: pET246-pUC57 12xMS2V6
FEATURES             Location/Qualifiers
     source          1..3578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..47)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     147..166
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     360..382
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     374..391
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     375..391
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1353..1369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1353..1369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1366..1388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1377..1393
                     /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)."
     promoter        complement(join(1401..1407,1408..1425,1426..1431))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1446..1467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1584..1601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1755..2343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1835..1854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2514..3305,3306..3374))
                     /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     3137..3156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3375..3479)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3547..3565
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
       61 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      121 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      181 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      241 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      301 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      361 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtacct cgcgaatgca
      421 tctagatcat ccaatctcga gatcgataga tcttccgtgt gagggtctct gtcgccagag
      481 ctcggtattc ctgagctcgc gtaacaatag gaatcccaga agtatccgca cgagtgctga
      541 tgatatatgc gcggtagtcc tgcgcaaatg gtcctgatga cgacgacaga gcacccatat
      601 tgaagtaatg caacggcagg gcgtattcgt ccctgtgaat gctcgcaagg cagatgcaat
      661 acacgcaccg ggggccgttt tggctcgcag ggtattcctc tgcgacagtg ctgccatagc
      721 agagtgtaaa ctttgcatgg gttggttgac agagaatggg gtattcctcc attggcaaaa
      781 aacttggcgg ccatctgcac accatgtatg atgaaggtag tcgagaagcg taatcgtctt
      841 ctcgtacatc ggcagaatac ctgcacaacc gaaaagatga atagaataga agtggtcggt
      901 agtcctgacc acacgagtag atttgactgt acgagtagac atgccgatat tctgcaccat
      961 gcatgcggta atcctgcatg acgtaagccg actccggtat ggtccaattt tgacggggaa
     1021 cagagtgtcc cacgcggtaa tcctgcgtgt cgattgttcg ggataatggt gcgatgcccc
     1081 gaaggtttcc tgactcgctt gtggggtagt cctccacatg tgacactggt gagcaatacg
     1141 gaatgttctt tgtagcaccg tggggccaca cagggtattc ctctgtgtcg ccctgggtga
     1201 cgttgctccc tgcgtatacc gcgcaagcga gagtgaagac gcggtagtcc tgcgtctccc
     1261 cgcgattgcc agggggctct gggatccatg attggatcgg atcccgggcc cgtcgactgc
     1321 agaggcctgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1381 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1441 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1501 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1561 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1621 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1681 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1741 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1801 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1861 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1921 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1981 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2041 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2101 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2161 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2221 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2281 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2341 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2401 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2461 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2521 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2581 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2641 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2701 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2761 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2821 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2881 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2941 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3001 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3061 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3121 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3181 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3241 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3301 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3361 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3421 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3481 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3541 tataaaaata ggcgtatcac gaggcccttt cgtctcgc
//
