LOCUS       sequence-482108-        6896 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Over-express E. coli Pol III subunit gamma-only in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6896)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 6896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Ec (gamma-only)-pET(11a)(ACH–)
FEATURES             Location/Qualifiers
     source          1..6896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..162)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        226..330
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(331..399,400..1191)
                     /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(549..568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1362..1950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1851..1870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2093..2110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2125..2264
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2190..2209)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2350..2372
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2366..2557)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3869..4951)
                     /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     4913..4932
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4952..5029)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5235..5254
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5338..5357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5338..5356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5357..5381
                     /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(6749..6767)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6763..6810
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
       61 tagcaattta actgtgataa actaccgcat taaagctggt cgatgataag ctgtcaaaca
      121 tgagaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
      181 gataataatg gtttcttagc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      301 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      361 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      421 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      481 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      541 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
      601 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      661 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      721 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      781 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      841 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
      901 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
      961 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1021 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1081 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1141 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1201 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1501 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1981 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2041 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2101 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2161 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2221 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2281 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2341 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2401 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     2461 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     2521 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     2581 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     2641 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     2701 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     2761 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     2821 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     2881 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     2941 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     3001 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc
     3061 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca
     3121 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt
     3181 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
     3241 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc
     3301 cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag
     3361 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
     3421 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
     3481 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc
     3541 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     3601 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     3661 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     3721 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     3781 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     3841 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3961 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4021 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     4081 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     4141 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     4201 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     4261 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     4321 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     4381 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     4441 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     4501 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     4561 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     4621 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     4681 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     4741 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     4801 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     4861 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     4921 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     4981 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     5041 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     5101 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     5161 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     5221 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     5281 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     5341 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ggagatatac
     5401 atatgagtta tcaggtctta gcccgaaaat ggcgcccaca aacctttgct gacgtcgtcg
     5461 gccaggaaca tgtgctgacc gcactggcga acggcttgtc gttagggcgt attcatcatg
     5521 cttatctttt ttccggcacc cgtggcgtcg gaaaaacctc tatcgcccga ctgctggcga
     5581 aggggctaaa ctgcgaaacc ggcattaccg cgacgccgtg cggcgtgtgc gataactgtc
     5641 gtgaaatcga gcaggggcgc tttgtcgatc tgattgaaat cgacgccgcc tcgcgcacca
     5701 aagttgaaga tacccgcgac ctgctggata acgtccagta cgctccggcg cgtggtcgtt
     5761 tcaaagttta tctgatcgac gaagtgcaca tgctgtcgcg ccacagcttt aacgcactgt
     5821 taaaaaccct tgaagagccg ccggagcacg ttaagtttct gctggcgacg accgatccac
     5881 agaaattgcc ggtgacgatt ttgtcacgct gtctgcaatt tcatctcaag gcgctggatg
     5941 tcgagcaaat tcgccatcag cttgagcaca tcctcaacga agaacatatc gctcacgagc
     6001 cgcgggcgct gcaattgctg gcacgcgccg ctgaaggcag cctgcgagat gccttaagtc
     6061 tgaccgacca ggcgattgcc agcggtgacg gccaggtttc aacccaggcg gtcagtgcga
     6121 tgctgggtac gcttgacgac gatcaggcgc tgtcgctggt tgaagcgatg gtcgaggcca
     6181 acggcgagcg cgtaatggcg ctgattaatg aagccgctgc ccgtggtatc gagtgggaag
     6241 cgttgctggt ggaaatgctc ggcctgttgc atcgtattgc gatggtacaa ctttcgcctg
     6301 ctgcacttgg caacgacatg gccgccatcg agctgcggat gcgtgaactg gcgcgcacca
     6361 taccgccgac ggatattcag ctttactatc agacgctgtt gattggtcgc aaagaattac
     6421 cgtatgcgcc ggaccgtcgc atgggcgttg agatgacgct gctgcgcgcg ctggcattcc
     6481 atccgcgtat gccgctgcct gagccagaag tgccacgaca gtcctttgca cccgtcgcgc
     6541 caacggcagt aatgacgcca acccaggtgc cgccgcaacc gcaatcagcg ccgcagcagg
     6601 caccgactgt accgctcccg gaaaccacca gccaggtgct ggcggcgcgc cagcagttgc
     6661 agcgcgtgca gggagcaacc aaagcgaaga aggaataagg atccggctgc taacaaagcc
     6721 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     6781 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc
     6841 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtg
//
