LOCUS       sequence-98503-e        8381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of Multibow set of constructs--each FP gene is initially not 
            expressed and then adopts a permanent ON or OFF status upon 
            Cre-mediated recombination. Encodes membrane bound mKO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8381)
  AUTHORS   Xiong F, Obholzer ND, Noche RR, Megason SG
  TITLE     Multibow: digital spectral barcodes for cell tracing.
  JOURNAL   PLoS One. 2015 May 26;10(5):e0127822. doi: 
            10.1371/journal.pone.0127822. eCollection 2015.
  PUBMED    26010570
REFERENCE   2  (bases 1 to 8381)
  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: "PLoS One.";
            date: "2015-05-26"; volume: "10(5):e0127822. doi"; pages: " 
            10.1371/journal.pone.0127822. eCollection 2015"
COMMENT     Sequence Label: pMTB-Multibow-mO
FEATURES             Location/Qualifiers
     source          1..8381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      353..941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     842..861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1095..1112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1229..1250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1265..1270,1271..1288,1289..1295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1319
                     /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     1308..1330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1327..1343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1327..1343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1362..1382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1364..1382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(1459..1470,1471..1658)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        4373..4391
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4373..4390
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(4446..4479)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(4487..4520)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(4530..4563)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4577..4586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5280..5401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5484..5517)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      5636..5645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    6532..6653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6668..6701)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(6755..6892,6893..6904)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6959..6976)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6959..6975)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6968..6990)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7117..7572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7204..7223)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7414..7435
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7598..7702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7703..7771,7772..8381,1..182)
                     /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(7921..7940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
       61 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      481 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1021 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1081 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1201 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1261 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1321 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     1381 gggaacaaaa gctggagctc caccgcggtg gcggcctagt ggatctgctg ggcttgctga
     1441 aggtaggggg tcaagaacca gaggtgtaaa gtacttgagt aattttactt gattactgta
     1501 cttaagtatt atttttgggg atttttactt tacttgagta caattaaaaa tcaatacttt
     1561 tacttttact taattacatt tttttagaaa aaaaagtact ttttactcct tacaatttta
     1621 tttacagtca aaaagtactt attttttgga gatcacttgg gcccggctcg agggcctagt
     1681 cattacctat actcaacgtt tactctgaga aacttccata ttttagaggt aaaatgctca
     1741 gaattgtaaa aagtcttgtc aaaatacttg gaatttgcca aaaacaaaat aaaagtcaca
     1801 tcacctcatt gtccttggta tcagcaagca cttttaacac aatcagctgt ttttgtttat
     1861 aactcccact gaaggccatg tgcattactc ccaagttaca cagtaaaaag tcacatgtag
     1921 tttatcatga tcactgctgc tcaagcacag ctttatctgt tattcagtgt ggcaatatct
     1981 gaaatgtgtg caaaatatta tttattgtag ctgatcacca acactttcag tgtttttttt
     2041 ttctactttt aatgagaatt atgttcttag gatttatcct tataaacctc tttaaaaaga
     2101 aatgttatgt ctgcatagac acgacatgaa attatgaaca gaaaatttaa ctatttaaga
     2161 tcaagacttg acatataaca tttctaatga acattaaacc tgaggtaggc ccacttgcaa
     2221 aaatgtaact gctatcactc actttataca tcattgtgta acttgtacac acaaaaatag
     2281 tgaagcaatg tggcatggaa tgcaggtccc attcattcaa agttattaaa gcactttctt
     2341 aaagtgcatt actactgagc taagaccagc tagttgctca tacaacatac tacaagtagt
     2401 gtcatcctaa tttggataaa aaaaatgtcc attttataca tggaaaagta ctcattagat
     2461 cccccattgt ttgtattacg gtatttcgtg aacacaagag gtaaatgacc tacagagctg
     2521 ctgttgtgtt agattgaaaa cacaacacag gatcatggag gcatccatca ttatgacgca
     2581 ttctgccact tgtaactctg cacagacttt ttgaaaagtt taattgagtc ttggcgttgt
     2641 cgcagaaaaa tacgaaaaat tctgtgttaa aatttgcaaa taaaaattcc agttttatag
     2701 gaaatcgcca tggccttgtt ctttaaccag ttaagcttcc ccttctttca ctctcaagtt
     2761 gcaagaagca agtgtagcaa tgtgcacgcg acagccgggt gtgtgacgct ggaccaatca
     2821 gagcacagag ctccgaaagt ttacctttta tggctagagc cgggtatgtg ccgtcatata
     2881 aaagagcgcg cccagctttt cagcctcact ttgagctcct ccacacgcag ctagtgcgga
     2941 atatcatctg cttgtaaccc attctcttaa gtcgacaacc ccccaaaccc aaggtgagtt
     3001 ggttttacag tttttaagcg tttaattata gtttctatct attaatgtta agtaaaatgt
     3061 gtaaatggat aaatatgatt tgatgaagtc tttagacagt tcataacggt ctctttgtag
     3121 ttatctacta ggtttgattt ataagtggtg gttcttttga taatctatgt tacatttaat
     3181 aaatattggc tgtatcttaa cttttttgta gcgttagtgg atgtggcagg tgagaatgtc
     3241 ggtaatttaa cgtgaccagt cgtaggcacg acattgaaca ttgaataggc cggtgtgaaa
     3301 tgggtgttaa gtcttaatat aaagttgtgt tctggctaat ctctctttaa cagccataaa
     3361 tgtgattttt aactacagtt ttgagggtgc tggtgagcat gttgcacact tgatggatag
     3421 ccggcatggg aagttctttg tgcaggcagt gctgcagcag ggtgtgacct actttagcta
     3481 gccggctaac cagctctcat ctgatgtaac gtaaacccca tcaatctaac atcggctcat
     3541 tcgtgcttta taagatcgaa gatcttgtta ttagtaacgg gtattggcat ttagttttaa
     3601 cttggttaat tttctttttg ttatgtgttc tatttaaatg tagcctgcag ccccctctgt
     3661 ctttaatgaa gctgaagatt aacgttaacg ttagtgaagt gttgtatccg ctatgaaaga
     3721 tttgaatttt agaggtgtgg tccactttaa taacggctgc ggagttctct ggctgcactt
     3781 cagccgctgt gactgtcgaa gcagagccca tcagtttttc aggatctcgg tgatttttga
     3841 ggctagacta tgaactgaac cgactgagag tgaccgtcca gaaatgcttt agtgtgaagt
     3901 ctccgctcac gaggagtccg tggactgact gcagatctgt gacgcagtaa cacccggcgg
     3961 cagacacccg ttgaaatgcg gttgtgtaat tgatcgcagg cgagggtgga agaggatgtg
     4021 aaacttcatt tgtgtaaaat ttagggagtg gtcctggtgt gatgaatgtc gaaatctgtt
     4081 cctttttact gagccctacg actctggctg agtgccacac cgccggcagc cgcaaagcgg
     4141 ctcaaaccat tgccttttat ggtaataatg agagaatgca gagggacttc ctttgtctgg
     4201 catatctgag gcgcgcgctg tcactagcgc ccaccagcgg tcagactgta gaatgcagag
     4261 caaaacagga agttgactcc agatggtcac atgcttgctg aaacgcctct ccctggcagc
     4321 agtgcaattc taacagtttc cctttctttt acagttcagc catcctaggc ctatttaggt
     4381 gacactatag aagagtacaa gctacttgtt ctttttgcag gatccgccac catgactagc
     4441 tcgagataac ttcgtatagt ataccttata cgaagttatc gagctcataa cttcgtatag
     4501 gatactttat acgaagttat aacgtcgaga taacttcgta tagcatacat tatacgaagt
     4561 tataccggca gatctcgcca ccatggtgag tgtgattaaa ccagagatga agatgaagta
     4621 cttcatggac ggctccgtca atgggcatga gttcacagtt gaaggtgaag gcacaggcaa
     4681 accttacgag ggacatcaag agatgacact acgcgtcaca atggccaagg gcgggccaat
     4741 gcctttctcg tttgacttag tgtcacacac gttctgttac ggccacagac cttttactaa
     4801 atatccagaa gagataccag actatttcaa acaagcattt cctgaaggcc tgtcatggga
     4861 aaggtcgttg cagttcgtgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4921 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4981 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     5041 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     5101 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaacacgtgc
     5161 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     5221 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5401 aaggcgtaaa ttgtaagcgt taatgctccg agggccgcca ccacctgttc ctgtaggaat
     5461 tggccgcgac tctaggatct aggataactt cgtataggat actttatacg aagttattga
     5521 tcaattcaca gccaccatgc tgtgctgcat cagaagaact aaaccggttg agaagaatga
     5581 agaggccgat caggagctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac
     5641 catgggagcg ccagctggag gtgcaggtgc agcaatggtg agcgtgatca aaccagagat
     5701 gaagatgcgc tactacatgg atggcagcgt gaacggccat gagttcacaa tcgagggcga
     5761 gggcacgggc cgcccttatg agggccatca ggagatgaca ctgcgcgtga caatggcaaa
     5821 aggcggccct atgccgttcg cattcgacct ggtgagccat gtgttctgct atggccatcg
     5881 ccctttcaca aagtaccctg aggagatccc tgactacttc aagcaggcat tccctgaggg
     5941 cctgagctgg gagcgcagcc tcgagttcga ggacggcggc agcgcaagcg tgagcgcaca
     6001 tatcagcctg cgcggcaaca cattctacca taagagcaaa ttcacgggcg tgaacttccc
     6061 tgcagacggc ccgatcatgc agaaccagag cgtggactgg gagcctagca cagagaaaat
     6121 cacagcaagc gacggcgtgc tgaagggcga tgtgacaatg tacctgaaac tggagggcgg
     6181 cggcaaccat aagtgccagt tcaaaacaac gtacaaggca gcgaagaaga tcctgaagat
     6241 gcctggcagc cattacatca gccatcgcct ggtgcgcaaa acagagggca acatcacaga
     6301 gctcgtggag gacgcagtgg cacataaagc agcagcatga taaatggtcc tgctggagtt
     6361 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     6421 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     6481 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     6541 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgcg
     6661 attcgagata acttcgtata gcatacatta tacgaagtta taccggaatt gtcgcgatcg
     6721 ccgaattatc ccgggtttaa acgcatgcag atctaatact caagtacaat tttaatggag
     6781 tactttttta cttttactca agtaagattc tagccagata cttttacttt taattgagta
     6841 aaattttccc taagtacttg tactttcact tgagtaaaat ttttgagtac tttttacacc
     6901 tctgtcaaga accatatgcc ggtacccaat tcgccctata ggcgtattac gcgcgctcac
     6961 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     7021 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     7081 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     7141 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     7201 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     7261 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     7321 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7381 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7441 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7501 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     7561 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7681 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7741 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7801 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7861 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7921 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7981 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     8041 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     8101 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     8161 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     8221 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     8281 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     8341 agttgcagga ccacttctgc gctcggccct tccggctggc t
//
