LOCUS       sequence-128176-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  AUTHORS   Huang L, Hung L, Odell M, Yokota H, Kim R, Kim SH
  TITLE     Structure-based experimental confirmation of biochemical function to
            a methyltransferase, MJ0882, from hyperthermophile Methanococcus 
            jannaschii.
  JOURNAL   J Struct Funct Genomics. 2002  . 2(3):121-7.
  PUBMED    12836702
REFERENCE   2  (bases 1 to 6001)
  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: "J Struct 
            Funct Genomics. 2002  . 2(3):121-7."
COMMENT     Sequence Label: pET21a.MJ0882
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             805..827
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    842..866
                     /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     complement(866..885)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(867..885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(969..988)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1194..1271
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1272..2354
                     /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(1291..1310)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3163..3354
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3348..3370)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3456..3598
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3511..3530
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3613..3630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3784..4372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3864..3883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4543..5334,5335..5403))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5166..5185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5404..5508)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5535..5990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5672..5693)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5884..5903
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tcctcataat ttcttacttt ttaagaccct ataacctcct
      241 tttattgtaa ctgtctcaac atttccaaag acatctttca tatatttagc taatgactta
      301 gccccttgct ttgtttgaat aactacccaa atctcgccat tatcttttaa aagttcttta
      361 ccttcctcaa taattctatg taaaacttcc tttccagctc ttattggtgg atttgttata
      421 atcttattat actttctgtc tttaacattt tcatataaat cgctatgaac taccctaata
      481 tcataattat ctaaattatt tagttttata ttctctttgg ctaattttat tgccctcctg
      541 tttatgtcag ccattgtagt tgatttaact tcatcagcta aggcaatgcc aataacacca
      601 taaccacagc ccaaatccaa gatgtcgtca tctttatcaa caactacgtt ttcaactaaa
      661 atttttgttc ctttatcaac ctttccataa gagaaaaccc cactatctgt tttaaatttt
      721 aatttttttc ctcttaaaat gtcttcaaca atttttacat ctgatttagt tgttggcttt
      781 tcagagaaat agtgcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
      841 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
      901 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
      961 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     1021 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     1081 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     1141 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     1201 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     1261 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     1321 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     1381 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     1441 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     1501 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     1561 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     1621 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     1681 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     1741 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     1801 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     1861 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     1921 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     1981 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     2041 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     2101 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     2161 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     2221 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     2281 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2341 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     2401 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     2461 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     2521 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     2581 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     2641 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg
     2701 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     2761 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     2821 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     2881 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
     2941 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
     3001 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
     3061 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
     3121 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
     3181 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
     3241 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
     3301 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3361 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3421 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     3481 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3541 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4201 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     4861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5521 aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     5581 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5641 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5701 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5761 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5821 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5881 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5941 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     6001 a
//
