LOCUS       sequence-405679-        5332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MTF1 protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5332)
  AUTHORS   Tang GQ, Paratkar S, Patel SS
  TITLE     Fluorescence mapping of the open complex of yeast mitochondrial RNA 
            polymerase.
  JOURNAL   J Biol Chem. 2009 Feb 27;284(9):5514-22. doi: 
            10.1074/jbc.M807880200. Epub 2008 Dec 30.
  PUBMED    19116203
REFERENCE   2  (bases 1 to 5332)
  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"; doi: 
            "10.1074/jbc.M807880200"; journalName: "J Biol Chem"; date: 
            "2009-02-27- 27"; volume: "284"; issue: "9"; pages: "5514-22"
COMMENT     Sequence Label: MTF1
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    733..873
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(799..818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     959..981
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1059..1136
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1137..2219
                     /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(1156..1175)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2451..2456,2457..2473,2474..2480)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2488..2504
                     /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     2493..2515
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    2521..2590
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             2641..2667
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             2683..2700
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3774..3791)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3944..4030
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4122..4149
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4168..4259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4260..4328,4329..5120)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4478..4497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5291..5332,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      781 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      841 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      901 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      961 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc
     1021 aattcgcgcg cgaaggcgaa gcggcatgca tttacgttga caccatcgaa tggtgcaaaa
     1081 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     1141 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     1201 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     1261 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     1321 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     1381 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     1441 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     1501 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     1561 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     1621 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     1681 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     1741 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     1801 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     1861 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     1921 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     1981 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     2041 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     2101 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     2161 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2221 cgcaacgcaa ttaatgtaag ttagcgcgaa ttgatctggt ttgacagctt atcatcgact
     2281 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     2341 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     2401 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     2461 atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2521 gcgccgctga gaaaaagcga agcggcactg ctctttaaca atttatcaga caatctgtgt
     2581 gggcactcga ccggaattat cgattaactt tattattaaa aattaaagag gtatatatta
     2641 atgtatcgat taaataagga ggaataaacc atggggggtt ctcatcatca tcatcatcat
     2701 ggtatggcta gctctgttcc aatccctggt attaaagata tctccaaact taagttcttt
     2761 tatggtttca aatatttatg gaatccaaca gtatacaata aaatctttga taagctggat
     2821 ttaactaaaa cgtataaaca tccagaagaa ttgaaagtac ttgacctcta ccctggcgtt
     2881 ggtatacaat cagctatttt ttataataaa tattgccccc gccaatattc tttactggaa
     2941 aaacgctcaa gtctctacaa gtttctgaat gcaaaattcg aagggtctcc attgcaaatt
     3001 ctaaaaagag atccatatga ctggtcaacc tactcgaatc taattgacga agagcgaata
     3061 tttgttcctg aagttcaatc ctcagatcat attaatgata aatttctaac cgttgctaat
     3121 gtgacggggg agggttctga aggtcttata atgcagtggc tgtcctgtat tggaaacaag
     3181 aactggttgt ataggtttgg taaagtaaag atgttattat ggatgccaag cacaacagct
     3241 agaaaacttc tagctaggcc aggcatgcat tctagatcca aatgttcagt agtaagggag
     3301 gcatttacag atactaaact cattgccata tcagacgcaa atgaattgaa gggatttgat
     3361 agtcaatgca tagaagaatg ggatcccatt ttatttagtg ctgcggaaat atggcctaca
     3421 aagggaaagc caattgcgtt agtagaaatg gacccaattg acttcgattt tgacgtagat
     3481 aattgggatt atgtcacgag gcacttaatg attttgaaaa gaacaccctt gaatactgtc
     3541 atggactcgc tgggacatgg tggccaacaa tattttaata gtagaatcac tgacaaagat
     3601 ctgctaaaaa agtgtccgat tgatttgaca aatgacgagt ttatatatct aacgaaatta
     3661 ttcatggagt ggcctttcaa accagatatt ttaatggatt ttgttgatat gtatcaaaca
     3721 gagcactctg gttgaaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     3781 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     3841 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     3901 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     3961 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     4021 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     4081 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     4141 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     4201 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4261 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4321 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4381 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4441 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4501 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4561 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4621 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4681 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4741 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4801 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4861 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4921 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4981 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5041 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5101 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5161 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5221 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5281 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa ac
//
