LOCUS       sequence-367105-        6424 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mCherry labeled with 8xPP7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6424)
  AUTHORS   Guo Y, Lee REC
  TITLE     Long-term imaging of individual mRNA molecules in living cells.
  JOURNAL   Cell Rep Methods. 2022 May 25;2(6):100226. doi: 
            10.1016/j.crmeth.2022.100226. eCollection 2022 Jun 20.
  PUBMED    35784652
REFERENCE   2  (bases 1 to 6424)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep 
            Methods."; date: "2022-05-25"; volume: "2(6):100226. doi"; pages: " 
            10.1016/j.crmeth.2022.100226. eCollection 2022 Jun 20"
COMMENT     Sequence Label: cmv-mCherry-8xPP7
FEATURES             Location/Qualifiers
     source          1..6424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(97..116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      910..1498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1399..1418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1652..1669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1786..1807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1822..1827,1828..1845,1846..1852)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1860..1876
                     /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     1865..1887
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1884..1900
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1884..1900
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1919..1939
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1921..1939
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(1990..2023)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            complement(2086..3442)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(2086..2283)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2289..3098)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3012..3031
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3099..3442)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      complement(3512..3699)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     3620..3644
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(3725..3758)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4228..4938)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4506..4525)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4775..4793
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      4935..4944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4966..5169)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4995..5015)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5170..5473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5530..5549)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5531..5549)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5559..5576)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5559..5575)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5568..5590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5717..6172
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5804..5823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6014..6035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6198..6302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6303..6371,6372..6424,1..739)
                     /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"
ORIGIN
        1 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
       61 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      121 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      181 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      241 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      301 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      361 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      421 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      481 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      541 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      601 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      661 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      721 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      781 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      841 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      901 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      961 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1021 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1081 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1141 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1201 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1261 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1321 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1381 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1441 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1501 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1561 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1621 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1681 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1741 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     1801 cactcattag gcaccccagg ctttacactt tatgctcccg gctcgtatgt tgtgtggaat
     1861 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
     1921 aattaaccct cactaaaggg aacaaaagct gggtaccgca taggccacta gtggatctga
     1981 tatcacctaa taacttcgta tagcatacat tatacgaagt tatattaagg gttctcgaga
     2041 gctcgttttc gacactggat ggcggcgtta gtatcgaatc gacagcagta tagcgaccag
     2101 cattcacata cgattgacgc atgatattac tttctgcgca cttaacttcg catctgggca
     2161 gatgatgtcg aggcgaaaaa aaatataaat cacgctaaca tttgattaaa atagaacaac
     2221 tacaatataa aaaaactata caaatgacaa gttcttgaaa acaagaatct ttttattgtc
     2281 agtactgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     2341 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2401 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2461 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     2521 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     2581 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     2641 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     2701 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     2761 tcaggatatt cttctaatac ctggaatgct gttttgccgg ggatcgcagt ggtgagtaac
     2821 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     2881 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     2941 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     3001 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     3061 aatcgcggcc tcgaaacgtg agtcttttcc ttacccatgg ttgtttatgt tcggatgtga
     3121 tgtgagaact gtatcctagc aagattttaa aaggaagtat atgaaagaag aacctcagtg
     3181 gcaaatccta accttttata tttctctaca ggggcgcggc gtggggacaa ttcaacgcgt
     3241 ctgtgagggg agcgtttccc tgctcgcagg tctgcagcga ggagccgtaa tttttgcttc
     3301 gcgccgtgcg gccatcaaaa tgtatggatg caaatgatta tacatgggga tgtatgggct
     3361 aaatgtacgg gcgacagtca catcatgccc ctgagctgcg cacgtcaaga ctgtcaagga
     3421 gggtattctg ggcctccatg tcgctggccg ggtgacccgg cggggacgag gcaagctaaa
     3481 cagatccacc ctgttatccc tagcggatct gccggtagag gtgtggtcaa taagagcgac
     3541 ctcatactat acctgagaaa gcaacctgac ctacaggaaa gagttactca agaataagaa
     3601 ttttcgtttt aaaacctaag agtcacttta aaatttgtat acacttattt tttttataac
     3661 ttatttaata ataaaaatca taaatcataa gaaattcgct tatttagaag tggcagatca
     3721 cctaataact tcgtatagca tacattatac gaagttatat taagggttag atctgccatt
     3781 tgtctcgagg tcgagaattc aaaaaaagca ccgactcggt gccacttttt caagttgata
     3841 acggactagc cttatttgaa cttgctactg ccatgtttga tccgtttggc tgctttggag
     3901 tgtttgccac ctttcggaca tttgagtagt ttgtgtgaca agcgaaccca tatgttctgc
     3961 ttgtcacacg tagcgatgcc atatcatctg ctactctact cctagcgatc ccatatgatc
     4021 tgctagttgt ccggaagcga agccatatct tctgcttctg gtggccgagc gaccccatat
     4081 ggtctgctcg tcactccgca gcgaggccat atcctctgct gctgcagccc cagcgagccc
     4141 atatgctctg ctggtcggat cggagcgacg ccatatcgtc tgctcctcat ggcagtagct
     4201 ctcaaacggg tcttcgtcct gcagggttta cttgtacagc tcgtccatgc cgccggtgga
     4261 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     4321 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
     4381 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
     4441 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     4501 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     4561 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     4621 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     4681 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     4741 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
     4801 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     4861 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
     4921 ctcgcccttg ctcaccatgg tggcgaccgg atccttcact aaacgagctc tgcttatata
     4981 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     5041 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     5101 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     5161 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     5221 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     5281 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     5341 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     5401 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     5461 aagttatgta acgcggaact ccactagtcc aaactggagc ggccgccacc gcggtggagc
     5521 tccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     5581 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5641 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5701 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5761 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5821 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5881 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5941 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6001 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6061 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6121 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     6181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6301 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6361 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6421 cacg
//
