LOCUS       sequence-334582-        4412 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 4412)
  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 4412)
  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: pET259-pUC57 24xMS2V6
FEATURES             Location/Qualifiers
     source          1..4412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2191..2207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2191..2207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2204..2226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2215..2231
                     /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(2239..2245,2246..2263,2264..2269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2284..2305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2422..2439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2593..3181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2673..2692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3352..4143,4144..4212))
                     /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     3975..3994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4213..4317)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4385..4403
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tcatccaatc tcgagatcga tagatcttcc gtgtgagggt ctctgtcgcc
      481 agagctcggt attcctgagc tcgcgtaaca ataggaatcc cagaagtatc cgcacgagtg
      541 ctgatgatat atgcgcggta gtcctgcgca aatggtcctg atgacgacga cagagcaccc
      601 atattgaagt aatgcaacgg cagggcgtat tcgtccctgt gaatgctcgc aaggcagatg
      661 caatacacgc accgggggcc gttttggctc gcagggtatt cctctgcgac agtgctgcca
      721 tagcagagtg taaactttgc atgggttggt tgacagagaa tggggtattc ctccattggc
      781 aaaaaacttg gcggccatct gcacaccatg tatgatgaag gtagtcgaga agcgtaatcg
      841 tcttctcgta catcggcaga atacctgcac aaccgaaaag atgaatagaa tagaagtggt
      901 cggtagtcct gaccacacga gtagatttga ctgtacgagt agacatgccg atattctgca
      961 ccatgcatgc ggtaatcctg catgacgtaa gccgactccg gtatggtcca attttgacgg
     1021 ggaacagagt gtcccacgcg gtaatcctgc gtgtcgattg ttcgggataa tggtgcgatg
     1081 ccccgaaggt ttcctgactc gcttgtgggg tagtcctcca catgtgacac tggtgagcaa
     1141 tacggaatgt tctttgtagc accgtggggc cacacagggt attcctctgt gtcgccctgg
     1201 gtgacgttgc tccctgcgta taccgcgcaa gcgagagtga agacgcggta gtcctgcgtc
     1261 tccccgcgat tgccaggggg ctctgggatc ttccgtgtga gggtctctgt cgccagagct
     1321 cggtattcct gagctcgcgt aacaatagga atcccagaag tatccgcacg agtgctgatg
     1381 atatatgcgc ggtagtcctg cgcaaatggt cctgatgacg acgacagagc acccatattg
     1441 aagtaatgca acggcagggc gtattcgtcc ctgtgaatgc tcgcaaggca gatgcaatac
     1501 acgcaccggg ggccgttttg gctcgcaggg tattcctctg cgacagtgct gccatagcag
     1561 agtgtaaact ttgcatgggt tggttgacag agaatggggt attcctccat tggcaaaaaa
     1621 cttggcggcc atctgcacac catgtatgat gaaggtagtc gagaagcgta atcgtcttct
     1681 cgtacatcgg cagaatacct gcacaaccga aaagatgaat agaatagaag tggtcggtag
     1741 tcctgaccac acgagtagat ttgactgtac gagtagacat gccgatattc tgcaccatgc
     1801 atgcggtaat cctgcatgac gtaagccgac tccggtatgg tccaattttg acggggaaca
     1861 gagtgtccca cgcggtaatc ctgcgtgtcg attgttcggg ataatggtgc gatgccccga
     1921 aggtttcctg actcgcttgt ggggtagtcc tccacatgtg acactggtga gcaatacgga
     1981 atgttctttg tagcaccgtg gggccacaca gggtattcct ctgtgtcgcc ctgggtgacg
     2041 ttgctccctg cgtataccgc gcaagcgaga gtgaagacgc ggtagtcctg cgtctccccg
     2101 cgattgccag ggggctctgg gatccatgat tggatcggat cccgggcccg tcgactgcag
     2161 aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2221 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2281 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2341 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2401 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2461 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2521 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2581 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2641 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2701 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2761 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2821 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2881 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2941 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3001 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3061 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3121 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3181 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3241 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3301 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3361 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3421 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3481 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3541 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3601 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3661 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3721 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3781 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3841 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3901 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3961 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4021 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4081 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4141 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4201 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4261 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4321 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4381 taaaaatagg cgtatcacga ggccctttcg tc
//
