LOCUS       sequence-156326-        5513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase expression under control of Pomc promoter (–646 to +65)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  AUTHORS   Kitamura T, Feng Y, Kitamura YI, Chua SC, Xu AW, Barsh GS, Rossetti 
            L, Accili D
  TITLE     Forkhead protein FoxO1 mediates Agrp-dependent effects of leptin on 
            food intake.
  JOURNAL   Nat Med. 2006 May . 12(5):534-40.
  PUBMED    16604086
REFERENCE   2  (bases 1 to 5513)
  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: "Nat Med. 
            2006 May . 12(5):534-40."
COMMENT     Sequence Label: Pomc-pGL3
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      777..786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             783..2435
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(849..867)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2476..2597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2487..2506)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2541..2560
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2845..2862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3016..3604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3096..3115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3775..4566,4567..4635))
                     /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     4398..4417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4636..4740)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4767..5222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4854..4873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5064..5085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5353..5401
                     /note="synthetic polyadenylation signal"
     misc_feature    5415..5506
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5455..5474
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagctg cacctcttgg ttaatagtct
       61 gaggctggca tgaccttgcc cttagagttt gggcacagaa ggacacctgt cttgaaataa
      121 gtattgggga atcaaggcag gcacacaccc cactccaaaa ggtagcctgc cttgggcggc
      181 cgtgactctt gacagcctct gttgtctccc ttctcaaaac ggaactgaga ttttggtttc
      241 acaagatatc acactttccc atcattgggg aaatctgcga cataacaaat ccccttcctc
      301 attagtgata tttacctcca aatgccagga aggcagatgg acgcacatag gtaattccac
      361 tccgatctgc aagatctcag aactaggcct gcctcgcaca gggacgctaa gcctctgtcc
      421 agttctaagt ggagattcaa caccattctt aattaagttc ttcctaacca ccagcgccag
      481 gtgtgcgctt cagcgggtct gtgttaacgc cagcctccgc gctttccagg cagatgtgcc
      541 ttgcgctcag ccaggaccgg gaagcccccc tcccgaggcc cgccgccccc cttcgctgca
      601 gaagcgctgc caggaaggtc acgtccaagg ctcacccacc caaccctgca agtataaaag
      661 aagagagaag agcgacaggg accaaacggg aggcgacgga agagaaaaga ggttaagagc
      721 agtgactaag agaggccact gaacatcaag cttggcattc cggtactgtt ggtaaagcca
      781 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
      841 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
      901 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
      961 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
     1021 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
     1081 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
     1141 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
     1201 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
     1261 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
     1321 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
     1381 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
     1441 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
     1501 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
     1561 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
     1621 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
     1681 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
     1741 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
     1801 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     1861 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     1921 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
     1981 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     2041 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
     2101 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
     2161 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
     2221 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     2281 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     2341 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     2401 taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc
     2461 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2521 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2581 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2641 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     2701 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     2761 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     2821 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2881 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3181 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     3241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3421 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4741 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     4801 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4861 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4921 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4981 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5041 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5101 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5161 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5221 ttcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5281 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     5341 tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     5401 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     5461 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg ata
//
