LOCUS       sequence-469257-        6510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  over-express E. coli PolX  (Pol III tau subunit - no gamma).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  AUTHORS   O'Donnell Lab
  TITLE     O'Donnell Lab Batch 1
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6510)
  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: 
            "unpublished"
COMMENT     Sequence Label: Ec dnaX(T-only)-pET(3c)
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      932..1520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1421..1440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1674..1691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1706..1845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1771..1790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1931..1953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1947..2138)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3596..3615
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3699..3718
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3699..3717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5738..5756)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5752..5799
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6165..6170,6171..6187,6188..6193))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6220..6238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6306..6410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6411..6479,6480..6510,1..761)
                     /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"
ORIGIN
        1 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
       61 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      121 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
      181 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      241 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      301 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      361 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      421 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
      481 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      541 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      601 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      661 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      721 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      781 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      841 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1321 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1561 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1621 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1681 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     1741 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     1801 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     1861 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     1921 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     1981 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     2041 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     2101 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     2161 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     2221 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     2281 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     2341 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     2401 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     2461 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     2521 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     2581 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     2641 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     2701 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     2761 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     2821 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     2881 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     2941 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     3001 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     3061 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     3121 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     3181 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     3241 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     3301 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     3361 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact
     3421 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     3481 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     3541 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     3601 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     3661 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggag
     3721 accacaacgg tttccctcta gaaggagata taaatatgag ttatcaggtc ttagcccgaa
     3781 aatggcgccc acaaaccttt gctgacgtcg tcggccagga acatgtgctg accgcactgg
     3841 cgaacggctt gtcgttaggg cgtattcatc atgcttatct tttttccggc acccgtggcg
     3901 tcggaaaaac ctctatcgcc cgactgctgg cgaaggggct aaactgcgaa accggcatta
     3961 ccgcgacgcc gtgcggcgtg tgcgataact gtcgtgaaat cgagcagggg cgctttgtcg
     4021 atctgattga aatcgacgcc gcctcgcgca ccaaagttga agatacccgc gacctgctgg
     4081 ataacgtcca gtacgctccg gcgcgtggtc gtttcaaagt ttatctgatc gacgaagtgc
     4141 acatgctgtc gcgccacagc tttaacgcac tgttaaaaac ccttgaagag ccgccggagc
     4201 acgttaagtt tctgctggcg acgaccgatc cacagaaatt gccggtgacg attttgtcac
     4261 gctgtctgca atttcatctc aaggcgctgg atgtcgagca aattcgccat cagcttgagc
     4321 acatcctcaa cgaagaacat atcgctcacg agccgcgggc gctgcaattg ctggcacgcg
     4381 ccgctgaagg cagcctgcga gatgccttaa gtctgaccga ccaggcgatt gccagcggtg
     4441 acggccaggt ttcaacccag gcggtcagtg cgatgctggg tacgcttgac gacgatcagg
     4501 cgctgtcgct ggttgaagcg atggtcgagg ccaacggcga gcgcgtaatg gcgctgatta
     4561 atgaagccgc tgcccgtggt atcgagtggg aagcgttgct ggtggaaatg ctcggcctgt
     4621 tgcatcgtat tgcgatggta caactttcgc ctgctgcact tggcaacgac atggccgcca
     4681 tcgagctgcg gatgcgtgaa ctggcgcgca ccataccgcc gacggatatt cagctttact
     4741 atcagacgct gttgattggt cgcaaagaat taccgtatgc gccggaccgt cgcatgggcg
     4801 ttgagatgac gctgctgcgc gcgctggcat tccatccgcg tatgccgctg cctgagccag
     4861 aagtgccacg acagtccttt gcacccgtcg cgccaacggc agtaatgacg ccaacccagg
     4921 tgccgccgca accgcaatca gcgccgcagc aggcaccgac tgtaccgctc ccggaaacca
     4981 ccagccaggt gctggcggcg cgccagcagt tgcagcgcgt gcagggagca accaaagcga
     5041 agaagagtga accggcagcc gctacccgcg cgcggccggt gaataacgct gcgctggaaa
     5101 gactggcttc ggtcaccgat cgcgttcagg cgcgtccggt gccatcggcg ctggaaaaag
     5161 cgccagccaa aaaagaagcg tatcgctgga aggcgaccac tccggtgatg cagcaaaaag
     5221 aagtggtcgc cacgccgaag gcgctgaaaa aagcgctgga acatgaaaaa acgccggaac
     5281 tggcggcgaa gctagcggca gaagccattg agcgcgaccc gtgggcggca caggtgagcc
     5341 aactttcgct accaaaactg gtcgaacagg tggcgttaaa tgcctggaaa gaggagagcg
     5401 acaacgcagt atgtctgcat ttgcgctcct ctcagcggca tttgaacaac cgcggtgcac
     5461 agcaaaaact ggctgaagcg ttgagcatgt taaaaggttc aacggttgaa ctgactatcg
     5521 ttgaagatga taatcccgcg gtgcgtacgc cgctggagtg gcgtcaggcg atatacgaag
     5581 aaaaacttgc gcaggcgcgc gagtccatta ttgcggataa taatattcag accctgcgtc
     5641 ggttcttcga tgcggagctg gatgaagaaa gtatccgccc catttgagga tccggctgct
     5701 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     5761 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     5821 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
     5881 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
     5941 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
     6001 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     6061 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     6121 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     6181 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     6241 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6301 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6361 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6421 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6481 acccagaaac gctggtgaaa gtaaaagatg
//
