LOCUS       sequence-169260-        6075 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 6075)
  AUTHORS   Kim JB, Sarraf P, Wright M, Yao KM, Mueller E, Solanes G, Lowell BB,
            Spiegelman BM
  TITLE     Nutritional and insulin regulation of fatty acid synthetase and 
            leptin gene expression through ADD1/SREBP1.
  JOURNAL   J Clin Invest. 1998 Jan 1. 101(1):1-9.
  PUBMED    9421459
REFERENCE   2  (bases 1 to 6075)
  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: "J Clin 
            Invest. 1998 Jan 1. 101(1):1-9."
COMMENT     Sequence Label: FAS promoter luciferase
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..106
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     98..115
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     99..115
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     122..141
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        122..140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    322..456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             741..2393
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(807..825)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2524..2589
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2719..2739
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3164..3298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3338..3356)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3339..3356)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3374..3390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3374..3390)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3387..3409)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3398..3414
                     /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(3422..3428,3429..3446,3447..3452))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3467..3488
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3605..3622)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3776..4364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3856..3875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4535..5326,5327..5395))
                     /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     5158..5177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5396..5500)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5578..6033)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5715..5736)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5927..5946
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
       61 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
      121 gtaatacgac tcactatagg gcgaattggg cccgacgtcg catgctcctc tagagcttgc
      181 agatctgcga ctctagagga tctgcgactc tagaggatca taatcagcca taccacattt
      241 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
      301 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
      361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      421 tccaaactca tcaatgtatc ttatcatgtc tggatccccg ggtaccgggg gctgcgcgtt
      481 ccttgtgctc cagcgcgcgc ctgtgcaggg tcccggctgg gggcggcgcg cgcgggcatc
      541 accccaccga cggcggcgcg ccgggtcccg gggcgcagcc ccgacgctca ttggcctggg
      601 cggcgcagcc aagctgtcag cccatgtggc gtggccgcgc ggggatggcc gcggtttaaa
      661 tagcgccggc gcggcctaga gggagccaga gagacggcag cagcgtcccg taagcttggc
      721 attccggtac tgttggtaaa atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
      781 tctatcctct agaggatgga accgctggag agcaactgca taaggctatg aagagatacg
      841 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtgaac atcacgtacg
      901 cggaatactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
      961 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     1021 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     1081 tgctcaacag tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc
     1141 aaaaaatttt gaacgtgcaa aaaaaattac caataatcca gaaaattatt atcatggatt
     1201 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     1261 ccggttttaa tgaatacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac
     1321 tgataatgaa ttcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa
     1381 ctgcctgcgt cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     1441 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     1501 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt
     1561 tacgatccct tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat
     1621 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     1681 ctgggggcgc acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc
     1741 cagggatacg acaaggatat gggctcactg agactacatc agctattctg attacacccg
     1801 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     1861 tggatctgga taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag
     1921 gacctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     1981 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     2041 tagttgaccg cttgaagtct ttaattaaat acaaaggata tcaggtggcc cccgctgaat
     2101 tggaatcgat attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg
     2161 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     2221 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     2281 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     2341 aaatcagaga gatcctcata aaggccaaga agggcggaaa gtccaaattg taaaatgtaa
     2401 ctgtattcag cgatgacgaa attcttagct attgtaatac tctagaggat ctttgtgaag
     2461 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct
     2521 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt
     2581 gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat
     2641 gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac
     2701 tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct
     2761 tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt
     2821 gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat
     2881 tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact
     2941 ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc
     3001 tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat
     3061 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3121 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     3301 gagagctccc aacgcgttgg atgcatagct tgagtattct atagtgtcac ctaaatagct
     3361 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3421 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3481 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3541 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3601 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3661 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3721 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3781 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3841 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3901 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3961 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4021 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4081 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4141 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4201 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4261 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4321 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4381 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4441 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4501 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4561 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4621 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4681 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4741 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4801 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4861 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4921 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4981 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5041 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5101 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5161 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5221 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5281 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5341 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5401 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5461 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5521 cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     5581 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5641 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5701 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5761 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5821 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5881 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5941 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6001 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     6061 ttgggaaggg cgatc
//
