LOCUS       sequence-366153-        6047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LECT2 V40 variant protein from T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6047)
  AUTHORS   Belonogov L, Taylor PE, Wong S, Morgan GJ
  TITLE     An amyloidosis-associated polymorphism does not alter LECT2 
            stability in vitro
  JOURNAL   bioRxiv 2022.03.01.482540
REFERENCE   2  (bases 1 to 6047)
  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: "bioRxiv 
            2022.03.01.482540"
COMMENT     Sequence Label: pET-11-LECT2-V40
FEATURES             Location/Qualifiers
     source          1..6047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     67..84
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    99..238
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(164..183)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     324..346
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(340..531)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1843..2925)
                     /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     2887..2906
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2926..3003)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3209..3228
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3312..3331
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3312..3330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3331..3355
                     /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             3370..3392
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3859..3877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3873..3920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4095..4100,4101..4117,4118..4123))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4150..4168)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4236..4340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4341..4409,4410..5201)
                     /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(4559..4578)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5372..5960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5861..5880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
       61 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
      121 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
      181 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
      241 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      301 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
      361 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
      421 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
      481 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
      541 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
      601 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
      661 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
      721 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
      781 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
      841 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
      901 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
      961 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     1021 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     1081 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     1141 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     1201 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     1261 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     1321 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc
     1381 tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc
     1441 ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg
     1501 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     1561 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     1621 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     1681 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     1741 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     1801 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     1861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1921 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     1981 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     2041 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     2101 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     2161 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     2221 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     2281 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2341 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2401 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2461 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2521 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     2581 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     2641 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     2701 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     2761 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     2821 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     2881 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     2941 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     3001 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     3061 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     3121 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     3181 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     3241 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     3301 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     3361 agaaataatt ttgtttaact ttaagaagga gatatacata tggggccatg ggctaatata
     3421 tgtgctggca agtcttccaa tgagatccgg acgtgtgacc gccatggctg tggacagtac
     3481 tctgctcaaa gaagtcagag gcctcaccag ggtgtggacg tcttgtgctc tgctggatct
     3541 actgtgtacg caccattcac tggaatgatt gtgggccagg agaaacctta tcaaaacaag
     3601 aatgctatca ataatggtgt tcgaatatct ggaagaggtt tttgtgtcaa aatgttctac
     3661 attaagccaa ttaagtataa aggtcctatt aagaagggag aaaaacttgg aactctattg
     3721 cccttgcaga aagtttatcc tggcatacaa tcgcatgtgc acattgaaaa ctgtgactcg
     3781 agtgacccta ctgcatacct gtagaatagg atccggctgc taacaaagcc cgaaaggaag
     3841 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3901 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
     3961 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
     4021 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     4081 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
     4141 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4201 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4261 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4321 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4381 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4441 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4501 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4561 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     4621 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4681 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4741 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4801 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4861 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     4921 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4981 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5041 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5101 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5161 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5221 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5581 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5641 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5701 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5761 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5821 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5881 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5941 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6001 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattct
//
