LOCUS       sequence-63666-e        8674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8674)
  AUTHORS   Phelps M, Stuelsatz P, Yablonka-Reuveni Z
  TITLE     Expression profile and overexpression outcome indicate a role for 
            betaKlotho in skeletal muscle fibro/adipogenesis.
  JOURNAL   FEBS J. 2016 May;283(9):1653-68. doi: 10.1111/febs.13682. Epub 2016 
            Apr 13.
  PUBMED    26881702
REFERENCE   2  (bases 1 to 8674)
  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: 
            "10.1111/febs.13682"; journalName: "FEBS J"; date: "2016-05"; 
            volume: "283"; issue: "9"; pages: "1653-68"
COMMENT     Sequence Label: pCMV-Kl-EGFP
FEATURES             Location/Qualifiers
     source          1..8674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     464..484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     558..577
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        558..576
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3663..3665,3666..3668,3669..4382)
                     /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(3663..3665,3666..3668,3669..4379)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3708..3729)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3969..3988)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4316..4337
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4427..4444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4433..4657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4789..5377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4869..4888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5706..5753
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5778..5797
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5985..6773)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6096..6115)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6700..6719
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6808..7165)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6822..6957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6974..7165
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7122..7142
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(7225..8016,8017..8085))
                     /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     7848..7867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8086..8190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8217..8672
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8304..8323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8514..8535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
       61 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      121 cgtcaatggg tggactattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      181 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      541 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      601 aacttaagct tagcatgcta gcccgcgccc ctcctcgccg cccgccgcgg ctggtgctgc
      661 tccgtttgct gttgctgcat ctgctgctgc tcgccctgcg cgcccgctgc ctgagcgctg
      721 agccgggtca gggcgcgcag acctgggctc gcttcgcgcg cgctcctgcc ccagaggccg
      781 ctggcctcct ccacgacacc ttccccgacg gtttcctctg ggcggtaggc agcgccgcct
      841 atcagaccga gggcggctgg cgacagcacg gcaaaggcgc gtccatctgg gacactttca
      901 cccatcactc tggggcggcc ccgtccgact ccccgatcgt cgtggcgccg tcgggtgccc
      961 cgtcgcctcc cctgtcctcc actggagatg tggccagcga tagttacaac aacgtctacc
     1021 gcgacacaga ggggctgcgc gaactggggg tcacccacta ccgcttctcc atatcgtggg
     1081 cgcgggtgct ccccaatggc accgcgggca ctcccaaccg cgaggggctg cgctactacc
     1141 ggcggctgct ggagcggctg cgggagctgg gcgtgcagcc ggtggttacc ctgtaccatt
     1201 gggacctgcc acagcgcctg caggacacct atggcggatg ggccaatcgc gccctggccg
     1261 accatttcag ggattatgcc gagctctgct tccgccactt cggtggtcag gtcaagtact
     1321 ggatcaccat tgacaacccc tacgtggtgg cctggcacgg gtatgccacc gggcgcctgg
     1381 ccccgggcgt gaggggcagc tccaggctcg ggtacctggt tgcccacaac ctacttttgg
     1441 ctcatgccaa agtctggcat ctctacaaca cctctttccg ccccacacag ggaggccggg
     1501 tgtctatcgc cttaagctcc cattggatca atcctcgaag aatgactgac tataatatca
     1561 gagaatgcca gaagtctctt gactttgtgc taggctggtt tgccaaaccc atatttattg
     1621 atggcgacta cccagagagt atgaagaaca acctctcgtc tcttctgcct gattttactg
     1681 aatctgagaa gaggctcatc agaggaactg ctgacttttt tgctctctcc ttcggaccaa
     1741 ccttgagctt tcagctattg gaccctaaca tgaagttccg ccaattggag tctcccaacc
     1801 tgaggcagct tctgtcttgg atagatctgg aatataacca ccctccaata tttattgtgg
     1861 aaaatggctg gtttgtctcg ggaaccacca aaagggatga tgccaaatat atgtattatc
     1921 tcaagaagtt cataatggaa accttaaaag caatcagact ggatggggtc gacgtcattg
     1981 ggtacaccgc gtggtcgctc atggacggtt tcgagtggca taggggctac agcatccggc
     2041 gaggactctt ctacgttgac tttctgagtc aggacaagga gctgttgcca aagtcttcgg
     2101 ccttgttcta ccaaaagctg atagaggaca atggctttcc tcctttacct gaaaaccagc
     2161 cccttgaagg gacatttccc tgtgactttg cttggggagt tgttgacaac tacgttcaag
     2221 tggacactac tctctctcag tttactgacc cgaatgtcta tctgtgggat gtgcatcaca
     2281 gtaagaggct tattaaagta gacggggttg tagccaagaa gagaaaacct tactgtgttg
     2341 atttctctgc catccggcct cagataacct tacttcgaga aatgcgggtc acccactttc
     2401 gcttctccct ggactgggcc ctgatcttgc ctctgggtaa ccagacccaa gtgaaccaca
     2461 cggttctgca cttctaccgc tgcatgatca gcgagctggt gcacgccaac atcactccag
     2521 tggtggccct gtggcagcca gcagccccgc accaaggcct gccacatgcc cttgcaaaac
     2581 atggggcctg ggagaacccg cacactgctc tggcgtttgc agactacgca aacctgtgtt
     2641 ttaaagagtt gggtcactgg gtcaatctct ggatcaccat gaacgagcca aacacacgga
     2701 acatgaccta tcgtgccggg caccacctcc tgagagccca tgccttggct tggcatctgt
     2761 acgatgacaa gtttagggcg gctcagaaag gcaaaatatc catcgccttg caggctgact
     2821 ggatagaacc ggcctgccct ttctctcaaa atgacaaaga agtggccgag agagttttgg
     2881 aatttgatat aggctggctg gcagagccta tttttggttc cggagattat ccacgtgtga
     2941 tgagggactg gctgaaccaa aaaaacaatt ttcttttgcc ctatttcacc gaagatgaaa
     3001 aaaagctagt ccggggttcc tttgacttcc tggcggtgag tcattacacc accattctgg
     3061 tagactggga aaaggaggat ccgatgaaat acaacgatta cttggaggta caggagatga
     3121 ctgacatcac atggctcaac tctcccagtc aggtggcagt ggtgccttgg gggctgcgca
     3181 aagtgctcaa ctggctaagg ttcaagtacg gagacctccc gatgtatgtg acagccaatg
     3241 gaatcgatga tgacccccac gccgagcaag actcactgag gatctattat attaagaatt
     3301 atgtgaatga ggctctgaaa gcctacgtgt tggacgacat caacctttgt ggctactttg
     3361 cgtattcact tagtgatcgc tcagctccca agtctggctt ttatcgatat gctgcgaatc
     3421 agtttgagcc caaaccatct atgaaacatt acaggaaaat tattgacagc aatggcttcc
     3481 tgggttctgg aacactggga aggttttgtc cagaagaata cactgtgtgc accgaatgtg
     3541 gattttttca aacccggaag tctttgctgg tcttcatctc gtttcttgtt tttactttta
     3601 ttatttctct tgctctcatt tttcactact ccaagaaagg ccagagaagt tataagaccg
     3661 gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3721 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3781 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3841 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3901 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3961 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4021 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4081 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     4141 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4201 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4261 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     4321 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     4381 aagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     4441 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4501 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4561 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4621 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagt
     4681 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4741 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4801 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4861 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4921 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4981 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5041 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5101 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5161 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5221 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5281 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5341 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5401 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5461 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5521 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     5581 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     5641 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     5701 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     5761 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     5821 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     5881 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     5941 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     6001 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     6061 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     6121 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     6181 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     6241 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     6301 tgatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     6361 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     6421 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     6481 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     6541 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     6601 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     6661 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     6721 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     6781 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
     6841 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     6901 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     6961 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     7021 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     7081 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     7141 ccacagctgg ttctttccgc ctcaggactc ttcctttttc aataaatcaa tctaaagtat
     7201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     8221 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     8281 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     8341 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     8401 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     8461 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     8521 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     8581 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     8641 gaattttaac aaaatattaa cgcttacaat ttac
//
