LOCUS       sequence-245685-        6538 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  AUTHORS   Istvan ES, Palnitkar M, Buchanan SK, Deisenhofer J
  TITLE     Crystal structure of the catalytic portion of human HMG-CoA 
            reductase: insights into regulation of activity and catalysis.
  JOURNAL   EMBO J. 2000 Mar 1. 19(5):819-30.
  PUBMED    10698924
REFERENCE   2  (bases 1 to 6538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J. 
            2000 Mar 1. 19(5):819-30."
COMMENT     Sequence Label: pGEXtev-HMG54kd
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             75..602
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(95..112)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(95..111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(104..126)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(704..709,710..725,726..732)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    740..756
                     /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     1390..1412
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1427..1447
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2276..2287
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3110..3132)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3276..3294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3362..3466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3467..3535,3536..4327)
                     /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(3685..3704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4498..5086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4987..5006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5240..5257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5330..5407
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5408..6490
                     /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(5427..5446)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
      121 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      181 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      241 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
      301 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
      361 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
      421 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
      481 aggaaggcca gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt
      541 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
      601 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
      661 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg
      721 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat
      781 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
      841 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
      901 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
      961 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
     1021 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
     1081 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
     1141 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
     1201 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
     1261 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
     1321 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
     1381 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catctcgaga atctttattt
     1441 tcagggcgcc atggcttcat cagtactggt gacacaggaa cctgaaattg aacttcccag
     1501 ggaacctcgg cctaatgaag aatgtctaca gatacttggg aatgcagaga aaggtgcaaa
     1561 attccttagt gatgctgaga tcatccagtt agtcaatgct aagcatatcc cagcctacaa
     1621 gttggaaact ctgatcgaaa ctcatgagcg tggtgtatct attcgccgac agttactttc
     1681 caagaagctt tcagaacctt cttctctcca gtacctacct tacagggatt ataattactc
     1741 cttggtgatg ggagcttgtt gtgagaatgt tattggatat atgcccatcc ctgttggagt
     1801 ggcaggaccc ctttgcttag atgaaaaaga atttcaggtt ccaatggcaa caacagaagg
     1861 ttgtcttgtg gccagcacca atagaggctg cagagcaata ggtcttggtg gaggtgccag
     1921 cagccgagtc cttgcagatg ggatgactcg tggcccagtt gtgcgtcttc cacgtgcttg
     1981 tgactctgca gaagtgaaag cctggctcga aacatctgaa gggttcgcag tgataaagga
     2041 ggcatttgac agcactagca gatttgcacg tctacagaaa cttcatacaa gtatagctgg
     2101 acgcaacctt tatatccgtt tccagtccag gtcaggggat gccatgggga tgaacatgat
     2161 ttcaaagggt acagagaaag cactttcaaa acttcacgag tatttccctg aaatgcagat
     2221 tctagccgtt agtggtaact attgtactga caagaaacct gctgctataa attggataga
     2281 gggaagagga aaatctgttg tttgtgaagc tgtcattcca gccaaggttg tcagagaagt
     2341 attaaagact accacagagg ctatgattga ggtcaacatt aacaagaatt tagtgggctc
     2401 tgccatggct gggagcatag gaggctacaa cgcccatgca gcaaacattg tcaccgccat
     2461 ctacattgcc tgtggacagg atgcagcaca gaatgttggt agttcaaact gtattacttt
     2521 aatggaagca agtggtccca caaatgaaga tttatatatc agctgcacca tgccatctat
     2581 agagatagga acggtgggtg gtgggaccaa cctactacct cagcaagcct gtttgcagat
     2641 gctaggtgtt caaggagcat gcaaagataa tcctggggaa aatgcccggc agcttgcccg
     2701 aattgtgtgt gggaccgtaa tggctgggga attgtcactt atggcagcat tggcagcagg
     2761 acatcttgtc aaaagtcaca tgattcacaa caggtcgaag atcaatttac aagacctcca
     2821 aggagcttgc accaagaaga cagcctgaat agcccgacag ttctgaactg gaacatgggc
     2881 attgggttct aaaggactaa cataaaatct gtgaattaaa aaagctcaat gcattgtctt
     2941 gtggaggatg aataaatgtg atcactgaga cagccacttg gtttttggct ctttcagaga
     3001 ggtctcaggt tctttccatg cagactcctc agatccccgg gtaccgagct cgaattcatc
     3061 gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3121 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3181 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     3241 gcgatagcgg agtgtaaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     3301 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3481 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3541 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3601 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3661 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     3721 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3781 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3841 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3901 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3961 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     4021 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4081 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4141 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4201 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4261 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4321 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4381 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4621 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5281 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     5341 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     5401 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     5461 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     5521 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     5581 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     5641 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     5701 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     5761 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     5821 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     5881 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     5941 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     6001 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     6061 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     6121 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     6181 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     6241 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     6301 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     6361 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     6421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6481 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggc
//
