LOCUS       sequence-351834-        7293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FBP17 c-terminal labeled with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7293)
  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 7293)
  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-FBP17
FEATURES             Location/Qualifiers
     source          1..7293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(3592..3594,3595..3597,3598..4311)
                     /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(3592..3594,3595..3597,3598..4308)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3637..3658)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3898..3917)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4245..4266
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4315..4334)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4380..4435
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4434..4453)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4796..4812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4796..4812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4809..4831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4820..4836
                     /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(4844..4850,4851..4868,4869..4874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4889..4910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4969..5164
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5015..5150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5170..5304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5207..5226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5261..5280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5372..5389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5543..6131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5623..5642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6302..7093,7094..7162))
                     /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     6925..6944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7163..7267)
                     /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 gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcaggtacc
     1741 atgagctggg gcaccgagct ctgggatcag tttgacaact tagaaaaaca cacacagtgg
     1801 ggaattgata ttcttgagaa atatatcaag tttgtgaaag aaaggacaga gattgaactc
     1861 agctatgcaa agcaactcag gaatctttca aagaagtacc aacctaaaaa gaactcgaag
     1921 gaggaagaag aatacaagta tacgtcatgt aaagctttca tttccaacct gaacgaaatg
     1981 aatgattacg cagggcagca tgaagttatc tccgagaaca tggcatcaca gatcattgtg
     2041 gacttggcac gctatgttca ggaactgaaa caggagagga aatcaaactt tcacgatggc
     2101 cgtaaagcac agcagcacat cgagacttgc tggaagcagc ttgaatctag taaaaggcga
     2161 tttgaacgcg attgcaaaga ggcggacagg gcgcagcagt actttgagaa aatggacgct
     2221 gacatcaatg tcacaaaagc ggatgttgaa aaggcccgac aacaagctca aatacgtcac
     2281 caaatggcag aggacagcaa agcagattac tcatccattc tccagaaatt caaccatgag
     2341 cagcatgaat attaccatac tcacatcccc aacatcttcc agaaaataca agagatggag
     2401 gaaaggagga ttgtgagaat gggagagtcc atgaagacat atgcagaggt tgatcggcag
     2461 gtgatcccaa tcattgggaa gtgcctggat ggaatagtaa aagcagccga atcaattgat
     2521 cagaaaaatg attcacagct ggtaatagaa gcttataaat cagggtttga gcctcctgga
     2581 gacattgaat ttgaggatta cactcagcca atgaagcgca ctgtgtcaga taacagcctt
     2641 tcaaattcca gaggagaagg caaaccagac ctcaaatttg gtggcaaatc caaaggaaag
     2701 ttatggccgt tcatcaaaaa aaataagctt atgtcccttt taacatcccc ccatcagcct
     2761 ccccctcccc ctcctgcctc tgcctcaccc tctgctgttc ccaacggccc ccagtctccc
     2821 aagcagcaaa aggaacccct ctcccaccgc ttcaacgagt tcatgacctc caaacccaaa
     2881 atccactgct tcaggagcct aaagcgtggg ggtgcaacac cggacgattt cagcaacctc
     2941 ccacctgaac aaagaaggaa aaagctgcag cagaaagtcg atgagttaaa taaagaaatt
     3001 cagaaggaga tggatcaaag agatgccata acaaaaatga aagatgtcta cctaaagaat
     3061 cctcagatgg gagacccagc cagtttggat cacaaattag cagaagtcag ccaaaatata
     3121 gagaaactgc gagtagagac ccagaaattt gaggcctggc tggctgaggt tgaaggccgg
     3181 ctcccagcac gcaacgagca ggcgcgccgg cagagcggac tgtacgacag ccagaaccca
     3241 cccacagtca acaactgcgc ccaggaccgt gagagcccag atggcagtta cacagaggag
     3301 cagagtcagg agagtgagat gaaggtgctg gccacggatt ttgacgacga gtttgatgat
     3361 gaggagcccc tccctgccat agggacgtgc aaagctctct acacatttga aggtcagaat
     3421 gaaggaacga tttccgtagt tgaaggagaa acattgtatg tcatagagga agacaaaggc
     3481 gatggctgga cccgcattcg gagaaatgaa gatgaagagg gttatgtccc cacttcatat
     3541 gtcgaagtct gtttggacaa aaatgccaaa gattcctgcg gccgcctcga gatggtgagc
     3601 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3661 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3721 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3781 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3841 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3901 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3961 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4021 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     4081 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     4141 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     4201 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     4261 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagatctttt
     4321 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     4381 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     4441 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     4501 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     4561 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     4621 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     4681 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     4741 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     4801 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4861 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     4981 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5041 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5101 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5161 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5221 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5281 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     5341 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5941 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6181 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6241 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6301 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6361 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6421 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6481 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6541 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6601 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6661 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6721 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6781 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6841 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6901 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6961 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7021 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7081 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7141 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7201 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7261 ttccgcgcac atttccccga aaagtgccac ctg
//
