LOCUS       sequence-123307-        5339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  provide MBS-PBS alternating sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  AUTHORS   Wu B, Chen J, Singer RH
  TITLE     Background free imaging of single mRNAs in live cells using split 
            fluorescent proteins.
  JOURNAL   Sci Rep. 2014 Jan 9;4:3615. doi: 10.1038/srep03615.
  PUBMED    24402470
REFERENCE   2  (bases 1 to 5339)
  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: "Sci Rep."; 
            date: "2014-01-9"; volume: "4:3615. doi"; pages: " 
            10.1038/srep03615"
COMMENT     Sequence Label: Pcr4-12xMBS-PBS
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1711..1730)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1712..1730)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1737..1754)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1737..1753)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1746..1768)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1891..2193
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2072..2091
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2542..3336
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2542..3336
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2596..2615)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3206..3225
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(3586..4377,4378..4446))
                     /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     4209..4228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4570..5158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5059..5078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5312..5329
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttcggcga
      301 gatctgatga accctggaat actggagcga gcgacgccat atcgtctgct cgtttgaaga
      361 ttcgacctgg aggcacggct gatgctcgtg ctttcttggc aataagtacc gtaggatctg
      421 atgaaccctg gaatactgga gcgagcgacg ccatatcgtc tgctcgtttg aagattcgac
      481 ctggaggcac ggctgatgct cgtgctttct tggcaataag taccgtagga tctgatgaac
      541 cctggaatac tggagcgagc gacgccatat cgtctgctcg tttgaagatt cgacctggag
      601 gcacggctga tgctcgtgct ttcttggcaa taagtaccgt aggatctgat gaaccctgga
      661 atactggagc gagcgacgcc atatcgtctg ctcgtttgaa gattcgacct ggaggcacgg
      721 ctgatgctcg tgctttcttg gcaataagta ccgtaggatc tgatgaaccc tggaatactg
      781 gagcgagcga cgccatatcg tctgctcgtt tgaagattcg acctggaggc acggctgatg
      841 ctcgtgcttt cttggcaata agtaccgtag gatctgatga accctggaat actggagcga
      901 gcgacgccat atcgtctgct cgtttgaaga ttcgacctgg aggcacggct gatgctcgtg
      961 ctttcttggc aataagtacc gtaggatctg atgaaccctg gaatactgga gcgagcgacg
     1021 ccatatcgtc tgctcgtttg aagattcgac ctggaggcac ggctgatgct cgtgctttct
     1081 tggcaataag taccgtagga tctgatgaac cctggaatac tggagcgagc gacgccatat
     1141 cgtctgctcg tttgaagatt cgacctggag gcacggctga tgctcgtgct ttcttggcaa
     1201 taagtaccgt aggatctgat gaaccctgga atactggagc gagcgacgcc atatcgtctg
     1261 ctcgtttgaa gattcgacct ggaggcacgg ctgatgctcg tgctttcttg gcaataagta
     1321 ccgtaggatc tgatgaaccc tggaatactg gagcgagcga cgccatatcg tctgctcgtt
     1381 tgaagattcg acctggaggc acggctgatg ctcgtgcttt cttggcaata agtaccgtag
     1441 gatctgatga accctggaat actggagcga gcgacgccat atcgtctgct cgtttgaaga
     1501 ttcgacctgg aggcacggct gatgctcgtg ctttcttggc aataagtacc gtaggatctg
     1561 atgaaccctg gaatactgga gcgagcgacg ccatatcgtc tgctcgtttg aagattcgac
     1621 ctggaggcac ggctgatgct cgtgctttct tggcaataag taccgtagga tcccgcgaag
     1681 ggcgaattcg cggccgctaa attcaattcg ccctatagtg agtcgtatta caattcactg
     1741 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1801 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1861 tcccaacagt tgcgcagcct atacgtacgg cagtttaagg tttacaccta taaaagagag
     1921 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc ggggcgacgg
     1981 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     2041 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     2101 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     2161 aacgccatta acctgatgtt ctggggaata taaatgtcag gcatgagatt atcaaaaagg
     2221 atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     2281 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     2341 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     2401 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     2461 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag
     2521 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2581 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2641 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2701 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     2761 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2821 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2881 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2941 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3061 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3301 atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta caatttcctg
     3361 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     3421 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3481 tccgctcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgac
     4501 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4561 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4621 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4681 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4741 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4801 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4861 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4921 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4981 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5041 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5101 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5161 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5221 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5281 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
