LOCUS       sequence-473456-        6153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of alpha-MSH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6153)
  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 6153)
  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- alpha-MSH
FEATURES             Location/Qualifiers
     source          1..6153
                     /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..809
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          810..1826
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1750..1772
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1834..1853
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1880..1904
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1905..1938)
                     /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(1971..2004)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2443..2476
                     /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    2509..2542
                     /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(2543..2567)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2577..3165
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2630..2650)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3048..3059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3195..3212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3201..3408
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3416..3556
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3416..3545
                     /label=AAV2 ITR
     rep_origin      3631..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3928..3949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4103..4122)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4222..4244
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4282..4300)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4368..4472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4473..4541,4542..5333)
                     /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(4691..4710)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5504..6092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5993..6012
                     /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 ggggcggcgc gggggggggg gcggggcgag gcggagaggt gcggcggcag
      721 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
      781 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
      841 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
      901 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac
      961 ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt
     1021 gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg
     1081 gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc
     1141 gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg
     1201 aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg
     1261 cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1321 cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1381 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1441 aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt
     1501 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg
     1561 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg
     1621 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc
     1681 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg
     1741 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
     1801 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1861 atcattttgg caaagaattg caagtttgta caaaaaagca ggctataact tcgtatagga
     1921 tactttatac gaagttatcc tcgagtaccg gtcacgcgta cgcaagcttg ataacttcgt
     1981 atagcataca ttatacgaag ttatcctagg gccacctcag cccaccggct tgccccagcg
     2041 gaagtgctcc atggagtagg agcgcttgcc ctcgcgtgga ctcggctctg gactgccatc
     2101 tccccacacc gcctcttcct ccgcacgcct ctgcgccgcg ctgccagcac tgctgctgtt
     2161 cctggggccg aagcggtccc agcggaagtg acccatgacg tacttccggg ggttttcagt
     2221 caggggctgt tcatctccgt tgccaggaaa cacgggcgtc tccagcgaga ggtcgagttt
     2281 gcaagcccgg atgcaagcca gcaggttgct ctccgtggtg aggtcctggc actggctgct
     2341 ctccaggcac cagctccaca catctatgga ggtctgaagc aggagggcca gcaacagggc
     2401 ccctgagcga ctgtagcaga atctcggcat ggtggctcta gaataacttc gtataaagta
     2461 tcctatacga agttataggt acctgagctc aacatcgata gcactagtat aacttcgtat
     2521 aatgtatgct atacgaagtt atacccagct ttcttgtaca aagtgggaat tccgataatc
     2581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2941 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     3001 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcgg gaattcctag
     3181 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3241 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3301 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3361 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagg gccgcaggaa
     3421 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3481 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3541 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3601 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3661 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3721 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3841 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3901 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3961 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4021 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4081 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4141 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4201 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4261 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4321 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4381 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4441 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4501 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4561 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4621 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4681 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4741 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4801 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4861 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4921 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4981 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5041 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5101 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5161 ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag
     5221 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5281 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5341 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5401 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5461 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5581 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5641 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5701 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5761 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5821 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5881 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5941 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6001 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6061 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6121 tcctggcctt ttgctggcct tttgctcaca tgt
//
