LOCUS       sequence-224336-        7151 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 7151)
  AUTHORS   Puigserver P, Wu Z, Park CW, Graves R, Wright M, Spiegelman BM
  TITLE     A cold-inducible coactivator of nuclear receptors linked to adaptive
            thermogenesis.
  JOURNAL   Cell 1998 Mar 20;92(6):829-39.
  PUBMED    9529258
REFERENCE   2  (bases 1 to 7151)
  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: "Cell"; 
            date: "1998-03-20"; volume: "92(6)"; pages: "829-39"
COMMENT     Sequence Label: GFP-PGC1
FEATURES             Location/Qualifiers
     source          1..7151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3879..4000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3916..3935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3970..3989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4007..4462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4144..4165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4356..4375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4489..4593
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4595..4952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4595..4786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4618..4638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4803..4938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4987..5781
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4987..5781
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5041..5060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5651..5670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5969..5988)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6013..6060
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6389..6977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6878..6897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgactcattc
     1321 gggagctgga tggcttggga catgtgcagc caagactctg tatggagtga catagagtgt
     1381 gctgctctgg ttggtgagga ccagcctctt tgcccagatc ttcctgaact tgacctttct
     1441 gaacttgatg tgaatgactt ggatacagac agctttctgg gtggattgaa gtggtgtagc
     1501 gaccaatcgg aaatcatatc caaccagtac aacaatgagc ctgcgaacat atttgagaag
     1561 atagatgaag agaatgaggc aaacttgcta gcggtcctca cagagacact ggacagtctc
     1621 cccgtggatg aagacggatt gccctcattt gatgcactga cagatggagc cgtgaccact
     1681 gacaacgagg ccagtccttc ctccatgcct gacggcaccc ctccccctca ggaggcagaa
     1741 gagccgtctc tacttaagaa gctcttactg gcaccagcca acactcagct cagctacaat
     1801 gaatgcagcg gtcttagcac tcagaaccat gcagcaaacc acacccacag gatcagaaca
     1861 aaccctgcca ttgttaagac cgagaattca tggagcaata aagcgaagag catttgtcaa
     1921 cagcaaaagc cacaaagacg tccctgctca gagcttctca agtatctgac cacaaacgat
     1981 gaccctcctc acaccaaacc cacagaaaac aggaacagca gcagagacaa atgtgcttcc
     2041 aaaaagaagt cccatacaca accgcagtcg caacatgctc aagccaaacc aacaacttta
     2101 tctcttcctc tgaccccaga gtcaccaaat gaccccaagg gttccccatt tgagaacaag
     2161 actattgagc gaaccttaag tgtggaactc tctggaactg caggcctaac tcctcccaca
     2221 actcctcctc ataaagccaa ccaagataac cctttcaagg cttcgccaaa gctgaagccc
     2281 tcttgcaaga ccgtggtgcc accgccaacc aagagggccc ggtacagtga gtgttctggt
     2341 acccaaggca gccactccac caagaaaggg cccgagcaat ctgagttgta cgcacaactc
     2401 agcaagtcct cagggctcag ccgaggacac gaggaaagga agactaaacg gcccagtctc
     2461 cggctgtttg gtgaccatga ctactgtcag tcactcaatt ccaaaacgga tatactcatt
     2521 aacatatcac aggagctcca agactctaga caactagact tcaaagatgc ctcctgtgac
     2581 tggcaggggc acatctgttc ttccacagat tcaggccagt gctacctgag agagactttg
     2641 gaggccagca agcaggtctc tccttgcagc accagaaaac agctccaaga ccaggaaatc
     2701 cgagcggagc tgaacaagca cttcggtcat ccctgtcaag ctgtgtttga cgacaaatca
     2761 gacaagacca gtgaactaag ggatggcgac ttcagtaatg aacaattctc caaactacct
     2821 gtgtttataa attcaggact agccatggat ggcctatttg atgacagtga agatgaaagt
     2881 gataaactga gctacccttg ggatggcacg cagccctatt cattgttcga tgtgtcgcct
     2941 tcttgctctt cctttaactc tccgtgtcga gactcagtgt caccaccgaa atccttattt
     3001 tctcaaagac cccaaaggat gcgctctcgt tcaagatcct tttctcgaca caggtcgtgt
     3061 tcccgatcac catattccag gtcaagatca aggtccccag gcagtagatc ctcttcaaga
     3121 tcctgttact actatgaatc aagccactac agacaccgca cacaccgcaa ttctcccttg
     3181 tatgtgagat cacgttcaag gtcaccctac agccgtaggc ccaggtacga cagctatgaa
     3241 gcctatgagc acgaaaggct caagagggat gaataccgca aagagcacga gaagcgggag
     3301 tctgaaaggg ccaaacagag agagaggcag aagcagaaag caattgaaga gcgccgtgtg
     3361 atttacgttg gtaaaatcag acctgacaca acgcggacag aattgagaga ccgctttgaa
     3421 gtttttggtg aaattgagga atgcaccgta aatctgcggg atgatggaga cagctatggt
     3481 ttcatcacct accgttacac ctgtgacgct ttcgctgctc ttgagaatgg atatacttta
     3541 cgcaggtcga acgaaactga cttcgagctg tacttttgtg gacggaagca atttttcaag
     3601 tctaactatg cagacctaga taccaactca gacgattttg accctgcttc caccaagagc
     3661 aagtatgact ctctggattt tgatagttta ctgaaggaag ctcagagaag cttgcgcagg
     3721 taacgtgttc ccaggctgag ggatgacagg gatccaccgg atctagataa ctgatcataa
     3781 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3841 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3961 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     4021 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4081 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4141 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4201 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4261 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4321 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4381 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4441 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4561 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4621 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4681 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4861 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4921 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4981 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5041 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5101 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5161 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5221 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5281 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5341 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5401 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5461 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5521 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5581 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5641 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5701 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5761 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5821 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5881 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5941 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6001 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6061 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6121 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6181 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6241 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6301 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6361 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6421 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6481 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6541 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6601 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6661 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6721 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6781 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6841 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6901 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6961 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7021 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7081 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     7141 atggagttcc g
//
