LOCUS       sequence-351808-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIP4 c-terminal labeled with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  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 7091)
  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-EGFP-CIP4
FEATURES             Location/Qualifiers
     source          1..7091
                     /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             join(3390..3392,3393..3395,3396..4109)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3390..3392,3393..3395,3396..4106)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3435..3456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3696..3715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4043..4064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4113..4132)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4178..4233
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4232..4251)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4594..4610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4594..4610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4607..4629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4618..4634
                     /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(4642..4648,4649..4666,4667..4672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4687..4708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4767..4962
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4813..4948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4968..5102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5005..5024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5059..5078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5170..5187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5341..5929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5421..5440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6100..6891,6892..6960))
                     /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     6723..6742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6961..7065)
                     /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 gkgcggcggc 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 gggcgaggag ctgttcaccg
     3421 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3481 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3541 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3601 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3661 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3721 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3781 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3841 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3901 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3961 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4021 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4081 ctctcggcat ggacgagctg tacaagtaaa gatctttttc cctctgccaa aaattatggg
     4141 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     4201 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     4261 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     4321 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     4381 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     4441 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     4501 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     4561 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4621 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4681 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4741 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     4801 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4861 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4921 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     4981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5101 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5161 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5221 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5281 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5341 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5401 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5461 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5521 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5581 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5641 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5701 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5761 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5821 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5881 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5941 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6001 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6061 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6181 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6301 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6421 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6661 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6721 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6781 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6841 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7021 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7081 agtgccacct g
//
