LOCUS       sequence-351835-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIP4 c-terminal labeled with mScarlet in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7070)
  AUTHORS   Taylor KL, Taylor RJ, Richters KE, Huynh B, Carrington J, McDermott 
            ME, Wilson RL, Dent EW
  TITLE     Opposing functions of F-BAR proteins in neuronal membrane 
            protrusion, tubule formation, and neurite outgrowth.
  JOURNAL   Life Sci Alliance. 2019 Jun 3;2(3). pii: 2/3/e201800288. doi: 
            10.26508/lsa.201800288. Print 2019 Jun.
  PUBMED    31160379
REFERENCE   2  (bases 1 to 7070)
  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: "Life Sci 
            Alliance."; date: "2019-06-3"; volume: "2(3). pii: 2/3/e201800288. 
            doi"; pages: " 10.26508/lsa.201800288. Print 2019 Jun"
COMMENT     Sequence Label: pCAX-mScarlet-CIP4
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3390..4085
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3523..3541)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3791..3810
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4092..4111)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4157..4212
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4211..4230)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4573..4589)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4573..4589)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4586..4608)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4597..4613
                     /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(4621..4627,4628..4645,4646..4651))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4666..4687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4746..4941
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4792..4927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4947..5081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4984..5003)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5038..5057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5149..5166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5320..5908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5400..5419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6079..6870,6871..6939))
                     /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     6702..6721
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6940..7044)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcaggtacca
     1741 tggattgggg cactgagctg tgggatcagt tcgaggtgct cgagcgccac acgcagtggg
     1801 ggctggacct gttggacaga tatgtaaagt tcgtgaaaga acgcaccgaa gtggaacagg
     1861 cttacgccaa acaactgcgg agcctggtga aaaaatatct gcccaagaga cctgccaagg
     1921 atgatcctga gtccaaattc agccagcaac agtccttcgt acagattctc caggaggtga
     1981 atgactttgc aggccagcgg gagctggtgg ctgagaacct cagtgtccgt gtatgtcttg
     2041 agctgaccaa gtactcacaa gagatgaaac aggagaggaa gatgcacttc caagaagggc
     2101 ggcgggccca gcagcagctg gaaaatggct ttaaacagct ggagaatagt aagcgtaaat
     2161 ttgagcggga ctgccgggag gcagagaagg cagcccagac tgctgaacgg ctagaccagg
     2221 atatcaacgc caccaaggct gatgtggaga aggccaagca gcaagcccac cttcggagtc
     2281 acatggccga agaaagcaaa aacgaatatg cggctcaact gcagcgcttc aaccgagacc
     2341 aagcccactt ctatttttca cagatgcccc agatattcga taagctccaa gacatggatg
     2401 aacgcagggc cacccgcctg ggtgccgggt atgggctcct gtcggaggcc gagctggagg
     2461 tggtgcccat aatagccaag tgcttggagg gcatgaaggt ggctgcaaat gctgtggatc
     2521 ccaagaacga ctcccacgtc cttatagagc tgcacaagtc aggttttgcc cgcccgggcg
     2581 acgtggaatt cgaggacttc agccagccca tgaaccgtgc accctccgac agcagtctgg
     2641 gcaccccctc ggatggacgg cctgaactcc gaggcccggg tcgcagccgc accaagcgct
     2701 ggccttttgg caagaagaac aagacagtgg tgaccgagga ttttagccac ttgcccccag
     2761 agcagcagcg aaaacggctt caacagcagt tggaagaacg cagtcgtgaa cttcagaagg
     2821 aggttgacca gagggaagcc ctaaagaaaa tgaaggatgt ctatgagaag acacctcaga
     2881 tgggggaccc cgccagcttg gagccccaga tcgctgaaac cctgagcaac attgaacggc
     2941 tgaaattgga agtgcagaag tatgaggcgt ggctggcaga agctgaaagt cgagtcctta
     3001 gcaaccgggg agacagcctg agccggcacg cccggcctcc cgaccccccc gctagcgccc
     3061 cgccagacag cagcagcaac agcgcatcac aggacaccaa ggagagctct gaagagcctc
     3121 cctcagaaga gagccaggac acccccattt acacggagtt tgatgaggat ttcgaggagg
     3181 aacccacatc ccccataggt cactgtgtgg ccatctacca ctttgaaggg tccagcgagg
     3241 gcactatctc tatggccgag ggtgaagacc tcagtcttat ggaagaagac aaaggggacg
     3301 gctggacccg ggtcaggcgg aaagagggag gcgagggcta cgtgcccacc tcctacctcc
     3361 gagtcacgct caattgcggc cgcctcgaga tggtgagcaa gggcgaggca gtgatcaagg
     3421 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     3481 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     3541 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     3601 gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     3661 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     3721 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     3781 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     3841 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     3901 gcggacgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     3961 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     4021 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     4081 acaagtaaag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     4141 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     4201 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     4261 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     4321 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4381 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4441 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4501 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt
     4561 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4621 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4681 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     4741 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4801 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4861 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4921 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     4981 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5041 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5161 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5221 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6181 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
