LOCUS       sequence-366027-        6966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse TIMP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  AUTHORS   Song T, Yao Y, Papoin J, Sherry B, Diamond B, Gu H, Blanc L, Zou YR
  TITLE     Host factor TIMP1 sustains long-lasting myeloid-biased hematopoiesis
            after severe infection.
  JOURNAL   J Exp Med. 2023 Dec 4;220(12):e20230018. doi: 10.1084/jem.20230018. 
            Epub 2023 Oct 18.
  PUBMED    37851372
REFERENCE   2  (bases 1 to 6966)
  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 Exp 
            Med."; date: "2023-12-4"; volume: "220(12):e20230018. doi"; pages: "
            10.1084/jem.20230018. Epub 2023 Oct 18"
COMMENT     Sequence Label: MSCV-TIMP1
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(455..1246,1247..1315))
                     /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     1078..1097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1316..1420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1488..1506
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1544..1566)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1666..1685
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1879..1901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2029..2048
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2248..2764
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2828..3169
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3236..3652
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3582..3604
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3620..3636
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        4324..4811
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4359..4382)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4744..4763
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      4829..4838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4835..4837,4838..4840,4841..5554)
                     /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             join(4835..4837,4838..4840,4841..5551)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4880..4901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5141..5160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5488..5509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5608..6122
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6284..6300
                     /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(6308..6314,6315..6332,6333..6338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6353..6374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6491..6508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6662..6966,1..284))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6742..6761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
       61 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1441 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1501 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1561 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1621 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1681 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1741 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1801 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1861 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     1921 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     1981 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2041 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2101 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     2161 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     2221 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     2281 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     2341 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     2401 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     2461 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     2521 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     2581 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     2641 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     2701 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
     2761 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc
     2821 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc
     2881 cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     2941 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     3001 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     3061 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     3121 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     3181 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     3241 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     3301 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     3361 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     3421 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     3481 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     3541 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     3601 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag
     3661 atctctcgag cagagcagat accatgatgg ccccctttgc atctctggca tctggcatcc
     3721 tcttgttgct atcactgata gcttccagta aggcctgtag ctgtgcccca ccccacccac
     3781 agacagcctt ctgcaactcg gacctggtca taagggctaa attcatgggt tccccagaaa
     3841 tcaacgagac caccttatac cagcgttata agatcaagat gactaagatg ctaaaaggat
     3901 tcaaggctgt gggaaatgcc gcagatatcc ggtacgccta caccccagtc atggaaagcc
     3961 tctgtggata tgcccacaag tcccagaacc gcagtgaaga gtttctcatc acgggccgcc
     4021 taaggaacgg aaatttgcac atcagtgcct gcagcttctt ggttccctgg cgtactctga
     4081 gccctgctca gcaaagagct ttctcaaaga cctatagtgc tggctgtggg gtgtgcacag
     4141 tgtttccctg tttatctatc ccttgcaaac tggagagtga cactcactgt ttgtggacgg
     4201 atcaggtcct cgtgggctct gaggactacc agagccgtca ctttgcttgc ctgccacgga
     4261 atccaggctt gtgcacctgg agatcccttg gggcccgatg acctgaagcc ttcgaattct
     4321 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     4381 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     4441 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     4501 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     4561 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     4621 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     4681 ggggtgggtc cgggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
     4741 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
     4801 cgggcctttc gacctgcagc ccaagcttgc caccatggtg agcaagggcg aggagctgtt
     4861 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4921 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4981 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     5041 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     5101 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     5161 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     5221 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     5281 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     5341 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     5401 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     5461 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     5521 gatcactctc ggcatggacg agctgtacaa gtaaagcggc catcgataaa ataaaagatt
     5581 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     5641 gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca
     5701 gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag
     5761 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca
     5821 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     5881 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     5941 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg
     6001 cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     6061 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     6121 atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa
     6181 gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag
     6241 ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat ccgctcacaa
     6301 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6361 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6421 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6481 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6541 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6601 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6661 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6721 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6781 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6841 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6901 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6961 actatc
//
