LOCUS       sequence-252122-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  generation of GST-tagged C-terminal part of U1-C protein using 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  TITLE     Tsubata CD72, SmB, and U1-C plasmids
REFERENCE   2  (bases 1 to 5175)
  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"
COMMENT     Sequence Label: pGEX-5X-1 U1-C-C
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(11..16,17..34,35..41)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    49..65
                     /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     73..89
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     73..89
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             85..612
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(105..122)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(105..121)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(114..136)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(714..719,720..735,736..742)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    750..766
                     /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             789..1442
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1400..1422
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1452..1463
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1756..1778)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1923..1941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2009..2113
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2114..2182,2183..2974)
                     /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(2332..2351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3145..3733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3634..3653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3887..3904
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3977..4054
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4055..5137
                     /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     complement(4074..4093)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
       61 aacaatttca cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta
      121 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      181 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      241 cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa
      301 agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag
      361 atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg
      421 ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa
      481 agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttggaat tagcttatcg
      541 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg
      601 tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
      661 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa
      721 ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      781 cagtattcat gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc
      841 gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag
      901 gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt
      961 atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca
     1021 agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag
     1081 cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc
     1141 tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat
     1201 gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg
     1261 ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt
     1321 gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt
     1381 atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa
     1441 aatcggatct gatcgaaggt cgtgggatcc ccgaattcat gcccgcaccc catatggggg
     1501 gccctcccat gatgccaatg atgggccctc ctcctcctgg gatgatgcca gtgggacctg
     1561 ctcctggaat gaggccgccc atgggaggcc atatgccaat gatgcctggg cccccaatga
     1621 tgagacctcc tgcccgtccc atgatggtgc ccactcggcc cggaatgact cgaccagaca
     1681 gataagtcga ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg
     1741 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1801 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1861 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa
     1921 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1981 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2041 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2101 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2161 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2221 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2281 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2341 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2401 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2461 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2521 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2581 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2641 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2701 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2761 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2821 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2881 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2941 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3001 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3061 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3121 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3181 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3241 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3301 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3361 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3421 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3481 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3541 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3601 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3661 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3721 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3781 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3841 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3901 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3961 acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     4021 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     4081 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     4141 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     4201 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     4261 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     4321 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     4381 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     4441 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     4501 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     4561 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4621 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4681 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4741 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     4801 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     4861 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     4921 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     4981 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     5041 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5101 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     5161 agctcactca ttagg
//
