LOCUS       sequence-403505-        7699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the LTB4-insensitive control sensor GEM-LTB4mut in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7699)
  AUTHORS   Tamas SX, Roux BT, Vamosi B, Dehne FG, Torok A, Fazekas L, Enyedi B
  TITLE     A genetically encoded sensor for visualizing leukotriene B4 
            gradients in vivo.
  JOURNAL   Nat Commun. 2023 Aug 1;14(1):4610. doi: 10.1038/s41467-023-40326-6.
  PUBMED    37528073
REFERENCE   2  (bases 1 to 7699)
  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: "Nat 
            Commun."; date: "2023-08-1"; volume: "14(1):4610. doi"; pages: " 
            10.1038/s41467-023-40326-6"
COMMENT     Sequence Label: pSB-CMV-MCS-Puro GEM-LTB4mut
FEATURES             Location/Qualifiers
     source          1..7699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      780..789
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     786..845
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1733..1754
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1860..1881)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2121..2140)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2755..2772)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2761..2985
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2992..3349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2992..3183
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3015..3035)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3200..3335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3377..3976
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3377..3396)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3873..3893
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3990..4009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    4019..4100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4044..4063
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4575..4595)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4575..4593)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4614..4630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4614..4630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4627..4649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4638..4654
                     /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(4662..4668,4669..4686,4687..4692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4707..4728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4845..4862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5016..5604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5096..5115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5775..6566,6567..6635))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6398..6417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6636..6740)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6767..7222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6854..6873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7064..7085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7348..7370
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7362..7379
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7363..7379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7389..7407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tgatgtcatg gctttagaag cttgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc
      721 ggatccacta gtccagtgtg gtggaattcc tgcagcccgg gggatccact agttctagcg
      781 ccaccatgga gacagacaca ctcctgctat gggtactgct gctctgggtt ccaggttcca
      841 ctggtgacac cagcctgtat aagaaggtgg gaaccaccgg taacactaca tcttctgccg
      901 cacccccctc actaggtgta gagttcatct ctctgctggc tatcatcctg ctgtcagtgg
      961 cgctggctgt ggggcttccc ggcaacagct ttgtggtgtg gagtatcctg aaaaggatgc
     1021 agaagcgctc tgtcactgcc ctgatggtgc tgaacctggc cctggccgac ctggccgtat
     1081 tgctcactgc tccctttttc cttcacttcc tggcccaagg cacctggagt tttggactgg
     1141 ctggttgccg cctgtgtcac tatgtctgcg gagtcagcat gtacgccagc gtcctgctta
     1201 tcacggccat gagtctagac cgctcactgg cggtggcccg cccctttgtg tcccagaagc
     1261 tacgcaccaa ggcgatggcc cggcgggtgc tggcaggcat ctgggtgttg tcctttctgc
     1321 tggccacacc cgtcctcgca tatgccacag tagtgccctg gaaaacgaac atgagcctgt
     1381 gcttcccgcg gtaccccagc gaagggcacc gggccttcca tctaatcttc gaggctgtca
     1441 cgggcttcct gctgcccttc ctggctgtgg tggccagcta ctcggacata gggcgtcggc
     1501 tgcaggcccg gcgcagcagc ctgattaacg tctatatcaa ggccgacaag cagaagaacg
     1561 gcatcaaggc gaacttccac atccgccaca acatcgagga cggcggcgtg cagctcgcct
     1621 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1681 acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc
     1741 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
     1801 gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1861 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg
     1921 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1981 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2041 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg
     2101 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2161 agggcgacac cctggtgaac cgcatcgagc ttaagggcat cgacttcaag gaggacggca
     2221 acatcctggg gcacaagctg gagtacaaca accacgatca gttccgccgc agtcgacgca
     2281 ccggccgcct ggtggtgctc atcatcctga ccttcgccgc cttctggctg ccctaccacg
     2341 tggtgaacct ggctgaggcg ggccgcgcgc tggccggcca ggccgccggg ttagggctcg
     2401 tggggaagcg gctgagcctg gcccgcaacg tgctcatcgc actcgccttc ctgagcagca
     2461 gcgtgaaccc cgtgctgtac gcgtgcgccg gcggcggcct gctgcgctcg gcgggcgtgg
     2521 gcttcgtcgc caagctgctg gagggcacgg gctccgaggc gtccagcacg cgccgcgggg
     2581 gcagcctggg ccagaccgct aggagcggcc ccgccgctct ggagcccggc ccttccgaga
     2641 gcctcactgc ctccagccct ctcaagttaa acgagctcaa ctaagaattc tgcagatatc
     2701 cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2761 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2821 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2881 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2941 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggaccgg actgaggcgg
     3001 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3061 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3181 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3241 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3301 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctagattt
     3361 tctgacccta gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3421 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3481 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3541 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3601 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3661 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3721 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtgtcgc ccgaccacca
     3781 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3841 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3901 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3961 caagcccggt gcctgaggat ctcgagctcg atgagtttgg acaaaccaca actagaatgc
     4021 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4081 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4141 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt acttaagagg
     4201 gatcgataag gatctagctt gtggaaggct actcgaaatg tttgacccaa gttaaacaat
     4261 ttaaaggcaa tgctaccaaa tactaattga gtgtatgtaa acttctgacc cactgggaat
     4321 gtgatgaaag aaataaaagc tgaaatgaat cattctctct actattattc tgatatttca
     4381 cattcttaaa ataaagtggt gatcctaact gacctaagac agggaatttt tactaggatt
     4441 aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg tatgtaaact
     4501 tccgacttca actgtatagg gatcctctag ctagagtcga cctcgagggg gggcccggta
     4561 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4621 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4681 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5161 accggatacc tgtccgcctt tctcccttcg ggaaccgtgg cgctttctca tagctcacgc
     5221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5761 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5821 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5881 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5941 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6001 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6061 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6121 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6181 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taacttggcc
     6241 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6301 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6361 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6421 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6481 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6541 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6601 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6661 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6721 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     6781 ggcgcattaa gcgcggcggg agtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6841 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6961 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     7021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7201 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     7261 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     7321 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     7381 agcgcgcgta atacgactca ctatagggcg aattggagct cggatcccta tacagttgaa
     7441 gtcggaagtt tacatacact taagttggag tcattaaaac tcgtttttca actactccac
     7501 aaatttcttg ttaacaaaca atagttttgg caagtcagtt aggacatcta ctttgtgcat
     7561 gacacaagtc atttttccaa caattgttta cagacagatt atttcactta taattcactg
     7621 tatcacaatt ccagtgggtc agaagtttac atacactaag ttgactgtgc ctttaaacag
     7681 cttggaaaat tccagaaaa
//
