LOCUS       sequence-346769-        6258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ism1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6258)
  AUTHORS   Jiang Z, Zhao M, Voilquin L, Jung Y, Aikio MA, Sahai T, Dou FY, 
            Roche AM, Carcamo-Orive I, Knowles JW, Wabitsch M, Appel EA, Maikawa
            CL, Camporez JP, Shulman GI, Tsai L, Rosen ED, Gardner CD, 
            Spiegelman BM, Svensson KJ
  TITLE     Isthmin-1 is an adipokine that promotes glucose uptake and improves 
            glucose tolerance and hepatic steatosis.
  JOURNAL   Cell Metab. 2021 Jul 28. pii: S1550-4131(21)00326-0. doi: 
            10.1016/j.cmet.2021.07.010.
  PUBMED    34348115
REFERENCE   2  (bases 1 to 6258)
  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 Metab.
            2021 Jul 28. pii: S1550-4131(21)00326-0. doi: 
            10.1016/j.cmet.2021.07.010."
COMMENT     Sequence Label: Mouse Ism1 with C-terminal Myc-6X-his tag
FEATURES             Location/Qualifiers
     source          1..6258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        158..349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     306..326
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(351..455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      482..937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(569..588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     779..800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1063..1085
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1077..1094
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1078..1094
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        1255..1634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1635..1838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1789..1809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1864..1883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1864..1882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3342..3371
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3390..3407
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3427..3443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3427..3443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3454..4076
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3653..3672)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4225..4813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4305..4324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5142..5189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5214..5233
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5421..6215)
                     /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             complement(5421..6215)
                     /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(5532..5551)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6142..6161
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6250..6258,1..349))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
       61 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      241 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      301 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
      361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
      421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
      481 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      541 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      601 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      661 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      781 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      841 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      901 aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca
      961 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     1021 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     1081 aaacgacggc cagtgccaag ctgatctata cattgaatca atattggcaa ttagccatat
     1141 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     1201 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt
     1261 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     1321 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     1381 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     1441 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1501 atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1561 atgcccagta catgacctta cgggactttc ctacttggca gtacatctac gtattagtca
     1621 tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg
     1681 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     1741 aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg
     1801 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagaatt
     1861 ttgtaatacg actcactata gggcggccgg gaattcgtcg actggatccg gtaccgagga
     1921 gatctgccgc cgcgatcgcc atggtgcgcc tggctgcgga actgctcctg ctgctggggc
     1981 tactgctgct cacgctgcac atcacggttc tgcgcggatc cggagcctcc gaccggcagg
     2041 acgcggccgc cggcaacgtc agcgggtccc agttgcagaa taacctcaac ctggagagtg
     2101 actccacgtc agaaaccagc tttcctctgt ccaaagaagc cccggaggag caccaggttg
     2161 tccaccaacc cttccccaga cagcgattcc ctccagagac tggacatcct tcactgcaga
     2221 gagacggccc cagatccttt ctccttgacc taccaaactt tccagatctt tccaaagctg
     2281 atatcaatgg gcagaatcca aacattcagg tcaccataga ggtggttgat ggccctgact
     2341 ccgaagcaga aaaagatcag catccagaga ataaacccag ctggtcgctc ccagctcccg
     2401 actggcgggc ctggtggcag aggtccttgt ccttggccag gacaaatagt ggggaccagg
     2461 atgacaagta tgacagtacc tcagatgaca gcaacttcct cagtgttcct agaggatggg
     2521 accgtccagc cccaggacac cggacttttg aaaccaaaga gcagccagag tatgattcca
     2581 cagacgggga gggtgattgg agtctctggt ctgtctgcag cgtcacctgt ggaaatggca
     2641 accagaaacg gacccgttct tgtggctatg catgcattgc cacagaatcc aggacctgtg
     2701 accgtccaaa ctgcccagga attgaagata ctttcaggac agcggccact gaagtgagtc
     2761 tgcttgcggg aagtgaggag tttaatgcca ccaagttgtt tgaagttgac atggacagct
     2821 gtgagcgatg gatgagctgc aaaagtgagt tcttaaagaa atacatgcac aaggtgatca
     2881 acgacctgcc cagctgcccc tgctcctacc ctactgaggt ggcctacagc acagctgaca
     2941 tctttgaccg catcaagcgc aaggacttcc gatggaagga tgctagtggg cccaaagaga
     3001 aactagagat ctacaagcct actgctcggt actgcatccg ctctatgctg tccctggaga
     3061 gtaccacact ggctgcccag cactgttgct atggtgacaa catgcagctc atcaccaggg
     3121 gcaaaggggc aggcactccc aatctcatca gcaccgagtt ctctgctgaa ctccactaca
     3181 aagtggatgt tctgccctgg attatctgca agggtgactg gagcagatat aatgaggccc
     3241 ggcctcccaa taacggacag aagtgcacag agagcccttc tgatgaggac tacatcaaac
     3301 agttccaaga agccagagag tacacgcgta cgcggccgct cgagcagaaa ctcatctcag
     3361 aagaggatct ggcagcaaat gatatcctgc accaccatca ccatcacgtt taaacggccg
     3421 gccgcggtca tagctgtttc ctgaacagat cccgggtggc atccctgtga cccctcccca
     3481 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     3541 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     3601 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     3661 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     3721 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     3781 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     3841 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     3901 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     3961 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     4021 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     4081 acgcggccag cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     4981 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     5041 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     5101 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     5161 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     5221 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     5281 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     5341 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     5401 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     5461 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     5521 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcgatc cgccacaccc
     5581 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     5641 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     5701 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     5761 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     5821 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     5881 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     5941 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     6001 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     6061 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     6121 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     6181 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     6241 tgatcgatct ttgcaaaa
//
