LOCUS       sequence-473451-        6647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of POMC gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6647)
  AUTHORS   Xu Y, Jiang Z, Li H, Cai J, Jiang Y, Otiz-Guzman J, Xu Y, Arenkiel 
            BR, Tong Q
  TITLE     Lateral septum as a melanocortin downstream site in obesity 
            development.
  JOURNAL   Cell Rep. 2023 May 30;42(5):112502. doi: 
            10.1016/j.celrep.2023.112502. Epub 2023 May 11.
  PUBMED    37171957
REFERENCE   2  (bases 1 to 6647)
  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 Rep.";
            date: "2023-05-30"; volume: "42(5):112502. doi"; pages: " 
            10.1016/j.celrep.2023.112502. Epub 2023 May 11"
COMMENT     Sequence Label: AAV-DIO-mPOMC
FEATURES             Location/Qualifiers
     source          1..6647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    148..168
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        553..830
                     /label=chicken beta-actin promoter
     intron          831..1847
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1771..1793
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1855..1874
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1901..1925
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1926..1959)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1992..2025)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(2038..2751,2752..2754,2755..2757))
                     /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             complement(join(2041..2751,2752..2754,2755..2757))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2083..2104)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2432..2451
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2691..2712
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2758..2814)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    3541..3574
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3607..3640
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3641..3665)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3689..3706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3695..3902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3910..4050
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3910..4039
                     /label=AAV2 ITR
     rep_origin      4125..4580
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4212..4231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4422..4443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4597..4616)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4716..4738
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4776..4794)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4862..4966
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4967..5035,5036..5827)
                     /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     complement(5185..5204)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5998..6586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6487..6506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttgct cgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
      601 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
      661 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
      721 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
      781 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      841 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      901 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      961 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
     1021 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
     1081 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
     1141 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     1201 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1261 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     1321 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     1381 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     1441 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     1501 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1561 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     1621 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     1681 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1741 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1801 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1861 tgctggttat tgtgctgtct catcattttg gcaaagaatt gcaagtttgt acaaaaaagc
     1921 aggctataac ttcgtatagg atactttata cgaagttatc ctcgagtacc ggtcacgcgt
     1981 acgcaagctt gataacttcg tatagcatac attatacgaa gttatcctag ggccacctta
     2041 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     2101 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
     2161 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     2221 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     2281 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     2341 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     2401 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     2461 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     2521 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
     2581 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     2641 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     2701 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatagg
     2761 cccggggttt tcttcaacat ctcctgcttg ctttaacaga gagaagttcg tggctccgct
     2821 tccctggccc ttcttgtgcg cgttcttgat gatggcgttc ttgaagagcg tcaccagggg
     2881 cgtctggctc ttctcggagg tcatgaagcc accgtaacgc ttgtccttgg gcgggttgct
     2941 ccagcggaag tgctccaccc ggtagggccc gtcgtccttc tccgcgtcgg actccaggac
     3001 ctgctccaag cctaatggcc gctcgccttc cagctccctc ttgaactcta ggggaaaggc
     3061 ctccgccgac tcgttctcag caacgttggg gtacaccttc accgggcgcc gtttcttgcc
     3121 caccggcttg ccccagcgga agtgctccat ggagtaggag cgcttgccct cgcgtggact
     3181 cggctctgga ctgccatctc cccacaccgc ctcttcctcc gcacgcctct gcgccgcgct
     3241 gccagcactg ctgctgttcc tggggccgaa gcggtcccag cggaagtgac ccatgacgta
     3301 cttccggggg ttttcagtca ggggctgttc atctccgttg ccaggaaaca cgggcgtctc
     3361 cagcgagagg tcgagtttgc aagcccggat gcaagccagc aggttgctct ccgtggtgag
     3421 gtcctggcac tggctgctct ccaggcacca gctccacaca tctatggagg tctgaagcag
     3481 gagggccagc aacagggccc ctgagcgact gtagcagaat ctcggcatgg tggctctaga
     3541 ataacttcgt ataaagtatc ctatacgaag ttataggtac ctgagctcaa catcgatagc
     3601 actagtataa cttcgtataa tgtatgctat acgaagttat acccagcttt cttgtacaaa
     3661 gtgggaattc ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3721 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3781 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3841 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     3901 gagggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3961 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4021 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4081 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4201 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4321 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4441 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4501 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4561 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     4621 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4681 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4741 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4801 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4981 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5041 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5101 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5161 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5221 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5281 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5341 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5401 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5461 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5521 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5581 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5641 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     5701 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5761 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5821 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5881 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5941 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6001 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6061 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6481 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6541 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6601 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
