LOCUS       sequence-252125-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  generation of GST-tagged U1-C protein using bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  TITLE     Tsubata CD72, SmB, and U1-C plasmids
REFERENCE   2  (bases 1 to 5448)
  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-FL
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(550..555,556..573,574..580)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    588..604
                     /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     612..628
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     612..628
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             624..1151
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(644..661)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(644..660)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(653..675)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1253..1258,1259..1274,1275..1281)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1289..1305
                     /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             1328..1981
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1939..1961
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1991..2002
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2568..2590)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2735..2753)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2821..2925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2926..2994,2995..3786)
                     /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(3144..3163)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3957..4545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4446..4465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4699..4716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4789..4866
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4867..5448,1..501)
                     /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(4886..4905)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
       61 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      121 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      181 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      241 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      301 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
      361 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
      421 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      481 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
      541 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      601 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
      661 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      721 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      781 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
      841 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
      901 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
      961 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1021 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga
     1081 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1141 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1201 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1261 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1321 agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
     1381 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
     1441 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
     1501 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
     1561 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
     1621 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
     1681 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
     1741 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
     1801 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
     1861 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
     1921 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
     1981 atcggatctg atcgaaggtc gtgggatccc cgaattcatg cccaagtttt attgtgacta
     2041 ctgcgataca tacctcaccc atgactctcc atctgtgaga aagacacact gcagtggaag
     2101 gaaacacaaa gagaatgtga aagactatta tcagaaatgg atggaagagc aggctcagag
     2161 cctgattgac aaaacaacgg ctgcatttca acaaggaaag atacctccta ctccattctc
     2221 tgctcctcct cctgcagggg cgatgatacc acctcccccc agccttccgg gtcctcctcg
     2281 ccctggtatg atgccagccc cccatatggg gggccctccc atgatgccaa tgatgggccc
     2341 tcctcctcct gggatgatgc cagtgggacc tgctcctgga atgaggccgc ccatgggagg
     2401 ccatatgcca atgatgcctg ggcccccaat gatgagacct cctgcccgtc ccatgatggt
     2461 gcccactcgg cccggaatga ctcgaccaga cagataagtc gactcgagcg gccgcatcgt
     2521 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2581 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2641 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     2701 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     2761 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2941 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3001 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3061 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3121 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     3181 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3241 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3301 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3361 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3421 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     3481 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3541 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3601 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3661 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3721 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3781 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3841 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3901 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4081 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4621 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4681 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4741 tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa
     4801 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     4861 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     4921 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     4981 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     5041 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     5101 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     5161 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     5221 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     5281 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     5341 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     5401 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcg
//
