LOCUS       sequence-197072-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Life imaging pCS2.0 Tag-RFP-T_UtrCH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  AUTHORS   Wu SK, Gomez GA, Michael M, Verma S, Cox HL, Lefevre JG, Parton RG, 
            Hamilton NA, Neufeld Z, Yap AS
  TITLE     Cortical F-actin stabilization generates apical-lateral patterns of 
            junctional contractility that integrate cells into epithelia.
  JOURNAL   Nat Cell Biol. 2014 Feb;16(2):167-78. doi: 10.1038/ncb2900. Epub 
            2014 Jan 12.
  PUBMED    24413434
REFERENCE   2  (bases 1 to 5623)
  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"; doi: "10.1038/ncb2900"; 
            journalName: "Nat Cell Biol"; date: "2014-02"; volume: "16"; issue: 
            "2"; pages: "167-78"
COMMENT     Sequence Label: pCS2.0 Tag-RFP-T-UtrCH
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..993
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1028..1046
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1028..1045
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1085..1816
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2668..2802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2692..2711)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2746..2765
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2915..2935)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2915..2933)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2954..2970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2954..2970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2967..2989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2978..2994
                     /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(3002..3008,3009..3026,3027..3032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3047..3068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3185..3202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3356..3944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3436..3455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4115..4906,4907..4975))
                     /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     4738..4757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4976..5080)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5106..5561)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5243..5264)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5455..5474
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
       61 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
      121 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
      181 cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
      241 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
      301 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
      361 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
      421 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
      481 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
      541 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
      601 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
      661 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
      721 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
      781 tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt
      841 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
      901 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
      961 gaggtctata taagcaatgc tcgtttaggg aaccgccatt ctgcctgggg acgtcggagc
     1021 aagcttgatt taggtgacac tatagaatac aagctacttg ttctttttgc aggatcccgc
     1081 caccatggtg tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat
     1141 ggagggcacc gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta
     1201 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
     1261 cgacatcctg gctaccagct tcatgtacgg cagcagaacc ttcatcaacc acacccaggg
     1321 catccccgac ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     1381 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct
     1441 catctacaac gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa
     1501 gaaaacactc ggctgggagg ccaacaccga gatgctgtac cccgctgacg gcggcctgga
     1561 aggcagaacc gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttcaa
     1621 gaccacatac agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt
     1681 ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga
     1741 ggtggctgtg gccagatact gcgacctccc tagcaaactg gggcacaaac ttaatggcat
     1801 ggacgagctg tacaagtccg gaaccatggc caagtatgga gaacatgaag ccagtcctga
     1861 caatgggcag aacgaattca gtgatatcat taagtccaga tctgatgaac acaatgacgt
     1921 acagaagaaa acctttacca aatggataaa tgctcgattt tcaaagagtg ggaaaccacc
     1981 catcaatgat atgttcacag acctcaaaga tggaaggaag ctattggatc ttctagaagg
     2041 cctcacagga acatcactgc caaaggaacg tggttccaca agggtacatg ccttaaataa
     2101 cgtcaacaga gtgctgcagg ttttacatca gaacaatgtg gaattagtga atataggggg
     2161 aaccgacatt gtggatggaa atcacaaact gactttgggg ttactttgga gcatcatttt
     2221 gcactggcag gtgaaagatg tcatgaagga tgtcatgtcg gacctgcagc agacgaacag
     2281 tgagaagatc ctgctcagct gggtgcgtca gaccaccagg ccctacagcc aagtcaacgt
     2341 cctcaacttc accaccagct ggacagatgg actcgccttt aatgctgtcc tccaccgaca
     2401 taaacctgat ctcttcagct gggataaagt tgtcaaaatg tcaccaattg agagacttga
     2461 acatgccttc agcaaggctc aaacttattt gggaattgaa aagctgttag atcctgaaga
     2521 tgttgccgtt cggcttcctg acaagaaatc cataattatg tatttaacat ctttgtttga
     2581 ggtgctacct cagcaagtca ccatagacta actcgagctc aagcttcgaa ttctgcagtc
     2641 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     2701 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2761 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2821 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     2881 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     2941 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3001 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3061 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3121 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3181 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3241 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3481 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3541 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3601 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3661 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3721 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3781 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3841 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3901 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3961 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4021 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4081 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4141 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4201 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4261 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4321 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4381 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4441 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4501 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4561 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4621 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4681 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4741 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4801 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4861 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4921 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4981 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5041 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5101 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5161 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5221 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5281 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5341 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5401 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5461 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5521 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     5581 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg cta
//
