LOCUS       sequence-380516-        7207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coding sequence of mouse ETNPPL is inserted in pET-15b vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  AUTHORS   Tsujioka H, Yamashita T
  TITLE     Utilization of ethanolamine phosphate phospholyase as a unique 
            astrocytic marker.
  JOURNAL   Front Cell Neurosci. 2023 Jan 30;17:1097512. doi: 
            10.3389/fncel.2023.1097512. eCollection 2023.
  PUBMED    36794261
REFERENCE   2  (bases 1 to 7207)
  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: "Front Cell 
            Neurosci."; date: "2023-01-30"; volume: "17:1097512. doi"; pages: " 
            10.3389/fncel.2023.1097512. eCollection 2023"
COMMENT     Sequence Label: pET-15b(ETNPPL)
FEATURES             Location/Qualifiers
     source          1..7207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..43)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        111..215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(216..284,285..1076)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(434..453)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1247..1835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1736..1755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1989..2006
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2021..2160
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2086..2105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2246..2268
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2262..2453)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3765..4847)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4809..4828
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4848..4925)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5131..5150
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5234..5253
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5234..5252
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5253..5277
                     /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)."
     RBS             5292..5314
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5333..5350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5360..5377
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6941..6959)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6955..7002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7177..7182,7183..7199,7200..7205))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 atgagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
       61 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      121 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      181 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      241 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      301 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      361 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      421 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
      481 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      541 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      601 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      661 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      721 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
      781 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      841 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      901 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      961 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1021 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1081 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1141 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1381 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1621 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1921 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1981 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     2041 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     2101 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     2161 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2221 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2281 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     2341 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     2401 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     2461 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     2521 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     2581 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     2641 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     2701 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     2761 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     2821 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     2881 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     2941 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     3001 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     3061 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     3121 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     3181 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg
     3241 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     3301 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     3361 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     3421 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     3481 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     3541 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     3601 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     3661 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     3721 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     3781 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3841 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     3901 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     3961 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     4021 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     4081 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4141 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     4201 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     4261 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     4321 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     4381 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     4441 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     4501 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     4561 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     4621 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     4681 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     4741 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     4801 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     4861 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     4921 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     4981 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     5041 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     5101 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5161 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     5221 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     5281 ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca
     5341 tcatcatcac agcagcggcc tggtgccgcg cggcagccat atgtgcgagc tctatagcaa
     5401 gcaggacacc ctggcgctga gggagaggca catcgggccc tcgtgcaaaa ttttctttgc
     5461 cgcggatccc atcaaaataa tgcgagccca ggggcagtac atgtttgatg agaaaggcga
     5521 acggtactta gactgcatca acaacgtggc gcatgtggga cactgtcacc cagaagtggt
     5581 caaagctgcc gcgaaacaga tggaattact aaatacaaat tctcgcttcc tccacgacaa
     5641 catcattgag tttgccaagc gccttactgc caccctgcct caggagctct ccgtttgcta
     5701 cttcacaaac tccggatccg aagccaatga cttagcctta cgtctggcac ggcagttccg
     5761 aggccaccag gatgtgatca cccttgacca tgcttaccac ggccacctgt cgtcattaat
     5821 tgagatcagt ccttataagt ttcagaaggg caaagatgtc aagagggaaa ctgtacatgt
     5881 ggcaccagct ccagacactt acagagggaa atacagagag gaccacgaag acccatccac
     5941 tgcatatgcc gacgaggtga agaagatcat tgaagaggct cacagcagcg gaaggaagat
     6001 tgctgccttt attgctgaat ccatgcagag ttgtggtgga caaattattc ctccagcagg
     6061 ctacttccag aaagtggctg aacacattca caaagcagga ggtgtgttca ttgctgacga
     6121 agttcaagta ggctttggca gagttgggag gtacttctgg agcttccaga tgtatggtga
     6181 agacttcgtt ccagacatcg tcaccatggg gaaacctatg ggcaacggtc acccgatatc
     6241 ctgtgtggtg acaaccaaag aaattgcaga agccttcagc agctcgggca tggaatattt
     6301 caatacgtat ggaggaaatc cagtgtcttg tgcagttggc ttggctgtgc tggatgtaat
     6361 tgaaaaggag aaccttcaag ggaatgctgt cagagtgggg acctatctca tggagctgct
     6421 gagtgaacag aaggctaagc atcccttgat aggggacatc agaggtgtcg gcctttttat
     6481 cggcatcgat ttggtgaagg accgtgagaa aagaacacct gcgacagctg aagcacagca
     6541 catcatctac gagatgaaag gaaagggggt acttctcagt gccgatggcc ctcacaggaa
     6601 cgtgctgaaa ataaaaccac ccatgtgctt cacggaagac gatgcaaagt tcctggtgga
     6661 ccaccttgac ggcattttaa cagttttaga agaagccatg gactccaaga gtggaactgt
     6721 gttctctgag aacacagctt acagaacaaa gatgcctaag gaaatacaag tggaattgcc
     6781 caaccttagt gccaccgagg ccagagaaat ccccagggga aagagaaatg gtgtatgctc
     6841 agaccagcaa gccctgctca gcaaaagact caagacatga atatgctcga ggatccggct
     6901 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     6961 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     7021 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     7081 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     7141 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     7201 gtcaaac
//
