LOCUS       sequence-145641-        8217 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Nt His-tag Equus caballus Sacsin 40-1317.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8217)
  TITLE     Sacsin | SIRPT1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pColdII-SIRPT1-Equus caballus
FEATURES             Location/Qualifiers
     source          1..8217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..81
                     /gene="cspA"
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    84..100
                     /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)."
     5'UTR           122..255
                     /gene="cspA"
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     CDS             256..270
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="MNHKV"
     CDS             271..288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     3'UTR           4178..4322
                     /gene="cspA"
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     primer_bind     complement(4363..4385)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4485..4504
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4521..4976)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4658..4679)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4870..4889
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5076..5180
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5181..5249,5250..6041)
                     /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(5399..5418)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6212..6800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6701..6720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6972..8054)
                     /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     8016..8035
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8055..8132)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat gttcggtcaa
      301 accactcctc ctctggttga cttcctgaag gacatcctgc gtcgttaccc agaaggtggt
      361 caaatcctga aggaactgat ccaaaacgct gaagacgcag gtgctactga agtaaagttc
      421 ctgtacgacg agactcagta cggtactgag actctgtggt ctaaggacat ggcacagtac
      481 cagggttctg ctctgtacgt atacaacaac gctgtcttca cccctgagga ctggcatggt
      541 atccaagaga tcgcacgttc tcgtaagaag gacgaccctc tgaaggtagg tcgtttcggt
      601 atcggtttca actctgtcta ccacatcacg gacgtccctt gtatcttctc cggtgaccaa
      661 atcggtatgc tggacccaca tcaaaccctg ttcggtcctc atgagtctgg tcaatgttgg
      721 aacctgaagg acgactctaa ggagatctct gagctgtctg accaattcgc acctttcatc
      781 ggtatcttcg gctccactaa ggagatgttc gtcaacggca acttcccggg tactttcttc
      841 cgcttccctc tgcgtctgca accatctcaa ctgtcttcta acctgtacaa caagcagaag
      901 gtcctggagc tgttcgagtc cttccgtgct gacgctgata ctgtactgct gttcctgaag
      961 tccgtccagg atgtatctct gcatgtccgt gaggcagatg gtactgagaa gctgatgttc
     1021 cgtgtaaccg catctgagtc tacggcactg aagcaggaac gtctgaactc tgtcaagatc
     1081 ctgggtactg caatctctaa ctactgcaag aagatcccga gcgactctgt gacttgcgtc
     1141 acttaccaca tcaacatcgt cctggaggac gagtctacta aggacacgca gaaaacctcc
     1201 tggctggtat gcaactccgt cggtggtcgt ggtgtatctt ctgcactgga ttctctggca
     1261 gatgagctga aattcgtgcc aatcatcggt atcgcaatgc cactgtctag ccgtgacgac
     1321 gaagagaaag gtgcaacttc tgacttctcc ggtaaagcgt tctgcttcct gccgctgcca
     1381 ccaggtgaag aatctaaaac tggtctgcca gtgcatatct ctggtttctt cggcctgact
     1441 gacaaccgtc gttctatcaa atggcgcgag ctggatcagt ggcgtgatcc agcagcactg
     1501 tggaatgaat tcctggtagt gaacgtggtt ccaaaagctt acgcaaccct gatcctggac
     1561 tctatcaaac gcctggagac tgagcgttct tctgacttcc cgttctctgt ggacgttatc
     1621 taccgtctgt ggccggatgt taacaaagtg cgtgtgcatt ggcagcgtgt tctggaaccg
     1681 ctgtttaacg aactgttcca gaacgcagtg ctgtatagca tcagcaacca ctgggtgcag
     1741 ctggaacagg catacttctc tgaactggac gactctctgg aatacaccga aaccgtgctg
     1801 aactacctgc agtcttctgg taaacagatc gtgaaagtgc cggctaacct ggctgccgcc
     1861 gtggaactgg ctgctccggg tgctgctcgt aaagttaccc cggcttgggt tcgtcaggta
     1921 ctgcgtaaat ctgctccgga aggtggtgct cgtgaacgtc tgcatctgct ggaatttgtt
     1981 ctgtctgatg aagcttattc tgaactgctg ggtctggaac tgctgccact gcagaatggc
     2041 cattttgttc cgttttctag cagcgtttct gaccaggacg ttgtgtacat cacctctgaa
     2101 gactacccgc gttccctgtt tccgggcctg gaaggccgtt ttattctgga taacctgaaa
     2161 ccgcatctgc tggctgctct gaaagaagcg gcgcagaccc gtggccgtcc gtgtacccaa
     2221 ctgcagctgc tgaatccgga acgttttgct cgtctgatta aagaagcgat gaatacgttt
     2281 tggccgggcc gtgaactgat tgttcagtgg tatccgtgtg ataaagataa acgccacccg
     2341 tccgtatcct ggctgaaaat ggtttggaaa aatctgtata tccacttcag cgaagacctg
     2401 accctgttcg atgaaatgcc gctgatcccg cgcaccgcgc tggaagaagg ccagatgtgt
     2461 gttgaactga ttcgcctgcg cattccgagc ctggttattc tggatgatga aagcgaagcg
     2521 cagctgccgg aatttctggc cgatattgtt cagaaactgg gcggcattat tctgaaaaaa
     2581 ctggatgcca gcattcagca cccgctgatt aaaaaataca tcaacagccc gctgccgtcc
     2641 gcggtactgc agattatgga aaaaatgccg ctgcagaaac tgtgcaacca gatcgcgagc
     2701 ctgctgccga cccacaaaga tgcgctgcgc aaatttctgg cgggcctgac cgatagctcc
     2761 gaaaaagaaa aacgcattat tcaggaactg acgatcttta aacgcgttaa ccactccagc
     2821 ggccagggcc tgtcctccta tacgaaactg aaaggctgta aagttctgca ccacaccgcg
     2881 aaactgccgc cgggcctgcg cctgagcgta ccggttattg attcctccga tgaagccacc
     2941 atccgcctgg cgaatatgct gaaaatcgaa accctgaaaa ccacctcctg tctgcagctg
     3001 gtactgaaag atatggaaaa tgccttttat agccacgaag aaattaccca tctgatgctg
     3061 tggattctgg aaaacctgtc cagcctgaaa aacgaaaacc cgaacgtgct ggactggctg
     3121 atgccgctga aattcatcca gatcagccag gaacagatcg tgtccgcgtc cgaactgttt
     3181 gatccggata ttgaaatcct gaaagacctg ttctacgacg aagaggagac ctgcttcccg
     3241 ccgagcgtgt tcaccagccc ggatattctg cactccctgc gccagattgg cctgaaaaat
     3301 gaagcgtccc tgaaagaaaa agacgtggtg caggtagcga aaaaaatcga aggcctgcag
     3361 gtgtcctcct gtccgaacca ggatgttctg ctgaaaaaag cgaaaacgct gctgctggtt
     3421 ctgaacaaaa accacgcgct gctgcagagc gccgaaggca aaatgaccct gaaaaaaatc
     3481 aaatgggttc cggcctgcaa agaacgcccg ccgaactatc cgggcagcct ggtttggaaa
     3541 ggcgatattt gcgatctgtg cgcgccgccg gatatgtgcg atgcgatgca cgccattctg
     3601 gttggcagct ccctgccgct ggtagaatcc gttcacatca acctggaaaa agcgctgggc
     3661 atcgttacca aaccgtccat tagcgcggtt ctgaaacact ttaaaatcgt tgtagattgg
     3721 tatacgtcca aaaccttttc cgatgaagat tactaccagt tccagcacat cctgctggaa
     3781 atctatggct tcatgcacga ccacctgaac gaaggcaaag acgccttccg cgccctgaaa
     3841 tttccgtggg tttggaccgg caaaaaattt tgcccgctga gccaggcggt tattaaaccg
     3901 ttccacgatc tggacctgca gccgtatctg cacagcgttc cgaaaaccat ggcgaaattc
     3961 caccagctgt tcaaagtttg cggcagcatt gaagaactga ccagcgacca catctccatg
     4021 gtgatccaga aagtttatct gaaatccgac caggacctga gcgaacagga aagcaaacag
     4081 aacctgcacc tgatgctgaa catcatccgc tggctgtatt ccaactaact cgagggatcc
     4141 gaattcaagc ttgtcgacct gcagtctaga taggtaatct ctgcttaaaa gcacagaatc
     4201 taagatccct gccatttggc ggggattttt ttatttgttt tcaggaaata aataatcgat
     4261 cgcgtaataa aatctattat tatttttgtg aagaataaat ttgggtgcaa tgagaatgcg
     4321 caggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4381 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4441 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4501 tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     4561 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4621 atcaaaagaa tagcccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4681 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4741 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4801 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4861 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4921 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
     4981 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5041 atcaggcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5101 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5161 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5221 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5281 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5341 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5401 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5461 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5521 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5581 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5641 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5701 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5761 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5821 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5881 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5941 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6001 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6061 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6121 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6181 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6241 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6301 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6361 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6421 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6481 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6541 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6601 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6661 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6721 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6781 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6841 gctggccttt tgctcacata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     6901 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     6961 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7021 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     7081 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     7141 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     7201 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     7261 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     7321 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     7381 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     7441 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     7501 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     7561 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     7621 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     7681 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     7741 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     7801 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     7861 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     7921 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     7981 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     8041 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     8101 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     8161 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgc
//
