LOCUS       sequence-154438-        7493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  AUTHORS   Peterson TR, Sengupta SS, Harris TE, Carmack AE, Kang SA, Balderas 
            E, Guertin DA, Madden KL, Carpenter AE, Finck BN, Sabatini DM
  TITLE     mTOR Complex 1 Regulates Lipin 1 Localization to Control the SREBP 
            Pathway.
  JOURNAL   Cell. 2011 Aug 5;146(3):408-20.
  PUBMED    21816276
REFERENCE   2  (bases 1 to 7493)
  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."; 
            date: "2011-08-5"; volume: "146(3)"; pages: "408-20"
COMMENT     Sequence Label: pRK5 FLAG wildtype, catalytic active lipin 1
FEATURES             Location/Qualifiers
     source          1..7493
                     /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      4029..4038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4042..4176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4079..4098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4133..4152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4245..4602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4245..4436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4268..4288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4453..4588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4622..4639)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4622..4638)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4653)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4851..5306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4938..4957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5148..5169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5323..5342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5442..5464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5502..5520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5588..5692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5693..5761,5762..6553)
                     /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(5911..5930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6724..7312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7213..7232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7466..7483
                     /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 cttgagcggc cgctaagtaa gtaaggatcc
     4021 ccagcttggc cgccatggcc caacttgttt attgcagctt ataatggtta caaataaagc
     4081 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4141 tccaaactca tcaatgtatc ttatcatgtc tggatcggga attaattcgg cgcagcacca
     4201 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     4261 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4321 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4381 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4441 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4501 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4561 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt aacagcttgg
     4621 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4681 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4741 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     4801 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4921 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4981 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5041 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     5101 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5161 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     5221 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5281 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     5341 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5401 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5461 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5521 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5581 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5641 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5701 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5761 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5821 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5881 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5941 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6001 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6061 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6121 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6181 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6241 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6301 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6361 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6421 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6481 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6541 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6601 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6661 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6721 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6781 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6841 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6901 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6961 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7021 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7081 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7141 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7201 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7261 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7321 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7381 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7441 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
