LOCUS       sequence-273222-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of c7orf59 and HBXIP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  AUTHORS   Shen K, Sabatini DM
  TITLE     Ragulator and SLC38A9 activate the Rag GTPases through noncanonical 
            GEF mechanisms.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Sep 18;115(38):9545-9550. doi: 
            10.1073/pnas.1811727115. Epub 2018 Sep 4.
  PUBMED    30181260
REFERENCE   2  (bases 1 to 4520)
  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: 
            "10.1073/pnas.1811727115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-09-18- 18"; volume: "115"; issue: "38"; pages: 
            "9545-9550"
COMMENT     Sequence Label: [pSK24] pACYCDuet His-c7orf59|HBXIP
FEATURES             Location/Qualifiers
     source          1..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             104..127
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        499..517
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    518..542
                     /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)."
     CDS             876..920
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(958..976)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      972..1019
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1240..1899)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1814..1833
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1900..2002)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2394..2411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2528..3073)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2608..2627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3298..4380)
                     /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     4342..4361
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4381..4458)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4504..4520,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agcctggaag ttctgttcca
      121 ggggcccgga tccatgactt ctgcgctgac ccaggggctg gagcgaatcc cagaccagct
      181 cggctacctg gtactgagtg aaggtgcagt gctggcgtca tctggggacc tggagaatga
      241 tgagcaggca gccagtgcca tctctgagct ggtcagcaca gcctgcggtt tccggctgca
      301 ccgcggcatg aatgtgccct tcaagcgcct gtctgtggtc tttggagaac acacactgct
      361 ggtgacggtg tcaggacaga gggtgtttgt ggtgaagagg cagaaccgag gtcgggagcc
      421 cattgatgtc tgagcggccg cataatgctt aagtcgaaca gaaagtaatc gtattgtaca
      481 cggccgcata atcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
      541 ccccatctta gtatattagt taagtataag aaggagatat acatatgatg gaggcgacct
      601 tggagcagca cttggaagac acaatgaaga atccctccat tgttggagtc ctgtgcacag
      661 attcacaagg acttaatctg ggttgccgcg ggaccctgtc agatgagcat gctggagtga
      721 tatctgttct agcccagcaa gcagctaagc taacctctga ccccactgat attcctgtgg
      781 tgtgtctaga atcagataat gggaacatta tgatccagaa acacgatggc atcacggtgg
      841 cagtgcacaa aatggcctct tgactcgagt ctggtaaaga aaccgctgct gcgaaatttg
      901 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
      961 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1021 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     1081 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     1141 cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca
     1201 accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc
     1261 atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc
     1321 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
     1381 catagtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
     1441 gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa
     1501 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
     1561 gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac
     1621 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
     1681 gaactccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
     1741 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
     1801 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
     1861 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
     1921 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     1981 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
     2041 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
     2101 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
     2161 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
     2221 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
     2281 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     2341 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     2401 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     2461 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     2521 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     2581 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     2641 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     2701 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     2761 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     2821 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     2881 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     2941 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3001 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3061 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     3121 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     3181 atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     3241 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3301 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3361 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     3421 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     3481 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     3541 tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc
     3601 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     3661 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     3721 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     3781 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     3841 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     3901 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     3961 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     4021 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     4081 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     4141 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     4201 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     4261 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     4321 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     4381 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     4441 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     4501 aattaatacg actcactata
//
