LOCUS       sequence-228172-        7589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ER/Golgi-tethered lipin-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  AUTHORS   Romani P, Brian I, Santinon G, Pocaterra A, Audano M, Pedretti S, 
            Mathieu S, Forcato M, Bicciato S, Manneville JB, Mitro N, Dupont S
  TITLE     Extracellular matrix mechanical cues regulate lipid metabolism 
            through Lipin-1 and SREBP.
  JOURNAL   Nat Cell Biol. 2019 Mar;21(3):338-347. doi: 
            10.1038/s41556-018-0270-5. Epub 2019 Feb 4.
  PUBMED    30718857
REFERENCE   2  (bases 1 to 7589)
  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"; doi: 
            "10.1038/s41556-018-0270-5"; journalName: "Nat Cell Biol"; date: 
            "2019-03"; volume: "21"; issue: "3"; pages: "338-347"
COMMENT     Sequence Label: pRK5 FLAG membrane-bound lipin-1 (TMD)
FEATURES             Location/Qualifiers
     source          1..7589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1173..1196
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4125..4134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4138..4272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4175..4194)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4229..4248
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4341..4698
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4341..4532
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4364..4384)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4549..4684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4718..4735)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4718..4734)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4727..4749)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4947..5402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5034..5053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5244..5265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5419..5438)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5538..5560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5598..5616)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5684..5788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5789..5857,5858..6649)
                     /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(6007..6026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6820..7408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7309..7328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7562..7579
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gattgaattc accatggaaa tggactacaa ggatgacgat gacaagggtg
     1201 gtggtggtgg tgcgtcgacg atgaattacg tggggcagct ggccggccag gtgtttgtga
     1261 cggtgaagga gctctacaag ggactgaacc ccgccacgct gtcgggatgc atcgacatca
     1321 ttgtcatccg gcagcccaat ggcagtctgc agtgctcccc tttccacgtc cgcttcggca
     1381 agatgggtgt cctccgctcc cgagagaaag tggtggacat agaaatcaat ggggagtccg
     1441 tggatttgca catgaagttg ggagacaacg gagaagcatt ttttgttcaa gagacggaca
     1501 acgatcagga aatcatcccc atgtacctgg ccacgtcccc catcctgtca gaaggagctg
     1561 cgagaatgga aagccagctg aagaggaact ctgtggacag aatcaggtgc ctggatccca
     1621 ctacagctgc ccagggcctg cctcccagcg acaccccatc cactggttct ctggggaaga
     1681 agagaaggaa aaggaggagg aaggcccagt tggacaatct caaaagagat gacaatgtca
     1741 actcatctga ggatgaggac atgtttccca tagagatgag ctcggatgag gacacagcac
     1801 cgatggatgg aagcagaact cttcctaatg atgtaccacc attccaagat gatattccta
     1861 aggaaaactt cccctcgatt tcaacgcacc cccagtcagc atcgtacccc agttcggaca
     1921 gagaatggtc ccccagcccc agcagcctgg tagattgcca gaggactccc cctcacctgg
     1981 ccgagggagt tctctctagc tcttgtcctc tgcagtcttg ccacttccat gcttcggaaa
     2041 gtccttcagg ctcccggccc tcaacaccaa aaagtgactc tgagctggtc agtaagtctg
     2101 cagacaggtt gacgccaaag aataacctgg aaatgctctg gctgtggggt gaattgccac
     2161 aggctgcaaa gtcctcttct ccacacaaga tgaaagagtc cagcccctta gggagccgga
     2221 agactcctga taaaatgaat tttcaggcca ttcacagcga gtcttcagat acttttagtg
     2281 accagtcgcc aacaatggcc cggggactgc tcatccacca gagtaaggcc cagacggaaa
     2341 tgcagtttgt gaacgaggag gatctcgagt ccttgggggc ggcagcccca ccttcacccg
     2401 tggccgaaga gctcaaggcc ccatatccca acaccgcaca gtcgtcgagc aagacagatt
     2461 ccccttccag gaagaaagat aaacggagcc gacaccttgg agctgatggt gtttatctgg
     2521 acgacctcac ggacatggac cctgaagtgg cagccctgta tttccccaag aatggggatc
     2581 ctggtgggct ccccaaacaa gccagtgaca acggagccag gtcagccaac cagtcaccac
     2641 agtcggtggg aggctcgggc atcgacagtg gtgtggagag cacctccgac agcctgaggg
     2701 acctgccatc catcgccatc tccctctgcg gtggcctcag tgaccacaga gagatcacca
     2761 aagatgcatt tttggaacaa gccgtgtcat atcagcaatt tgccgacaac cctgctatca
     2821 tcgatgaccc caacctcgtg gtcaaggttg gcaataagta ttacaactgg acaacagcag
     2881 ctcctctact tctggcgatg caggctttcc agaaaccttt gccaaaggcc actgtggaat
     2941 ccatcatgag agataagatg cccaaaaagg gaggaagatg gtggttttcc tggagaggaa
     3001 gaaatgccac aatcaaagag gaaagcaagc ctgaacagtg cctgactggg aaaggccaca
     3061 ataccggaga gcagcctgcc cagcttggcc tggccaccag gataaagcat gagtcatcct
     3121 ccagtgatga agagcacgca gccgccaagc catcaggttc gagccacctc tctctcttgt
     3181 ccaacgtcag ctacaaaaag accctgcggc tcacgtcgga gcagctgaaa agcttgaagt
     3241 tgaagaacgg ccccaatgat gtggtgttca gtgtcactac ccagtaccag ggcacctgtc
     3301 gctgcgaggg caccatctac ctgtggaatt gggacgacaa agtcatcatc tcagatatcg
     3361 atgggaccat cacaaggtct gatactcttg gtcacatttt gcccacgctg ggaaaggatt
     3421 ggactcacca gggcattgca aagctgtacc acaaagtaag ccagaatggc tacaagtttc
     3481 tctattgttc ggcacgtgcc attgggatgg cggacatgac gaggggctac ctgcactggg
     3541 tcaacgagag gggcacggtg cttccacagg gcccgcttct gctcagcccg agcagcctct
     3601 tctccgcctt gcacagagaa gtgattgaaa agaagccaga aaagttcaaa gtccagtgtt
     3661 tgacagacat caagaacctg tttttcccaa acacagaacc cttctatgct gcttttggga
     3721 accggcctgc tgatgtgtat tcctacaagc aagtgggagt gtccctgaat aggatcttca
     3781 ctgtgaaccc caagggtgag ctggtgcagg agcatgccaa gaccaacatc agctcgtacg
     3841 tgcggctctg cgaagtggtc gatcacgtct tcccattgct aaagagaagc cattcctgtg
     3901 acttcccctg ttcagacact ttcagtaact tcaccttttg gagagagcca ctgccacctt
     3961 ttgaaaacca ggacatgcat tcagcctcag ctggtggtgg tggtggtgcg acctccaacc
     4021 ggtggctcat ggtcaaaatc ttcctcatcc tcattgtctt cttcatcatc tttgtggtct
     4081 tccttgcttg agcggccgct aagtaagtaa ggatccccag cttggccgcc atggcccaac
     4141 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4201 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4261 catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga
     4321 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4381 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4441 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4561 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4621 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4681 ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc
     4741 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4801 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4861 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4921 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4981 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5041 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5101 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5161 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5221 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5281 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5341 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5401 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5461 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5521 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5581 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5641 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5701 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5761 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5821 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5881 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5941 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6001 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6061 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6121 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6181 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6241 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6301 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6361 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6421 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6481 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6541 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6601 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6661 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6721 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6781 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6841 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6901 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6961 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7021 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7081 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7141 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7201 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7261 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7321 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7381 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7441 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7501 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     7561 cagcgagtca gtgagcgagg aagcggaag
//
