LOCUS       sequence-490852-        4247 bp    DNA     circular SYN 28-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4247)
  AUTHORS   Bostrom P, Wu J, Jedrychowski MP, Korde A, Ye L, Lo JC, Rasbach KA, 
            Bostrom EA, Choi JH, Long JZ, Kajimura S, Zingaretti MC, Vind BF, Tu
            H, Cinti S, Hojlund K, Gygi SP, Spiegelman BM
  TITLE     A PGC1-alpha-dependent myokine that drives brown-fat-like 
            development of white fat and thermogenesis.
  JOURNAL   Nature. 2012 Jan 11;481(7382):463-8. doi: 10.1038/nature10777.
  PUBMED    22237023
REFERENCE   2  (bases 1 to 4247)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2012-01-11- 11"; volume: "481"; issue: "7382"; 
            pages: "463-8"
COMMENT     Sequence Label: pCR-blunt-TOPO-FNDC5
FEATURES             Location/Qualifiers
     source          1..4247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1135..1154)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1136..1154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1161..1178)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1161..1177)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1170..1192)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1315..1617
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1496..1515
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1966..2760
                     /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             1966..2760
                     /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(2020..2039)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2630..2649
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2960..2969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2966..3340
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3478..4066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3967..3986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4220..4237
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttcgta atatcaatca ccgtcagaag
      361 gtcctctcgc attctctact gtcttctgag ctgctcagag caagcactgt aaatgtgtca
      421 tcatatcttg ctgcggagga gacccccacc ctgatgctcc ggtgtgctgg tttctgatgc
      481 gctcttggtt ttctccttgt tgttattggg ctcgttgtcc ttgatgatat catactggcg
      541 gcagaagaga gctataacac ctgcccacat gaagaggacc acaacaatga tcagcacctc
      601 ccccgttcgc agctgctggt tcctccccat ctccttcatg gtcacctcat ctttgttctt
      661 tgaggccatc ttttcagcct cgcgtggggt cttgaagagc acaggctcac tggctgggct
      721 ctgtccctgg atggagatgg cctgcacatg gacgatatat tctgtgtcct cctccaggtc
      781 ccagagagcg caggaccggg tggtggtgtt cacctcctga atgaaccgga gcatccgcac
      841 atccttcttc tgctgagaga tggcaaagcc aatgaccact tcatcctcca ggacatccca
      901 gctgaccacg gcagagttgg ccttgaggtg ccggacggtc acgttcacag gggctgaggg
      961 gctgtccgcc tgcaccagcg cgaagcagac gcagcctagc cacaggcgga gcgcggcgcg
     1021 gggcggccag gcgcacggcc ctgggggcat ggcgactcct gggccaaggg cgaattctgc
     1081 agatatccat cacactggcg gccgctcgag catgcatcta gagggcccaa ttcgccctat
     1141 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1201 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1261 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt acggcagttt
     1321 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     1381 attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     1441 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     1501 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
     1561 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     1621 tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta gaaagccagt
     1681 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     1741 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     1801 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     1861 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     1921 aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     1981 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2041 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2101 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2161 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2221 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2281 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2341 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2401 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2461 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     2521 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2581 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2641 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     2701 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     2761 attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2821 tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2881 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2941 ataatagcac gtgaggaggg ccaccatggc caagttgacc agtgccgttc cggtgctcac
     3001 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3061 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3121 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     3181 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     3241 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     3301 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctaa aacttcattt
     3361 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3421 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3481 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3541 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3961 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4021 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4081 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4141 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4201 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
