LOCUS       sequence-469249-        5187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  over-express E. coli holC (Pol III chi subunit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  AUTHORS   O'Donnell Lab
  TITLE     O'Donnell Lab Batch 1
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5187)
  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            "unpublished"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 holC-pET(3c)
FEATURES             Location/Qualifiers
     source          1..5187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     344..361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    376..515
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(441..460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     601..623
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(617..808)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2266..2285
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2369..2388
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2369..2387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2419..2441
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3085..3103)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3099..3146
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3512..3517,3518..3534,3535..3540))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3567..3585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3653..3757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3758..3826,3827..4618)
                     /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(3976..3995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4789..5187,1..190)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
       61 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      361 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      421 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
      481 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
      541 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
      601 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
      661 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
      721 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
      781 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
      841 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
      901 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
      961 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     1021 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     1081 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     1141 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     1201 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     1261 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     1321 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     1381 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     1441 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     1501 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     1561 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     1621 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     1681 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     1741 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     1801 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     1861 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     1921 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     1981 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     2041 tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact cctgcattag
     2101 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     2161 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     2221 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     2281 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     2341 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg
     2401 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat gaaaaacgcg
     2461 acgttctacc ttctggacaa tgacaccacc gtcgatggct taagcgccgt tgagcaactg
     2521 gtgtgtgaaa ttgccgcaga acgttggcgc agcggtaagc gcgtgctcat cgcctgtgaa
     2581 gatgaaaagc aggcttaccg gctggatgaa gccctgtggg cgcgtccggc agaaagcttt
     2641 gttccgcata atttagcggg agaaggaccg cgcggcggtg caccggtgga gatcgcctgg
     2701 ccgcaaaagc gtagcagcag ccggcgcgat atattgatta gtctgcgaac aagctttgca
     2761 gattttgcca ccgctttcac agaagtggta gacttcgttc cttatgaaga ttctctgaaa
     2821 caactggcgc gcgaacgcta taaagcctac cgcgtggctg gtttcaacct gaatacggca
     2881 acctggaaat aatggaaaag acatataacc cacaagatat cgaacagccg ctttacgagc
     2941 actgggaaaa gcagggctac tttaagccta atggcgatga aagccaggaa agtttctgca
     3001 tcatgatccc gccgccgaac gtcaccggca gtttggatcc ggctgctaac aaagcccgaa
     3061 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3121 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg
     3181 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg
     3241 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc
     3301 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg
     3361 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
     3421 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
     3481 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
     3541 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3601 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3721 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3781 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3841 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3901 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3961 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     4021 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4081 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4141 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4201 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4261 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     4321 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4381 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4441 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4501 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4561 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4621 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4681 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4741 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4801 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4861 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4921 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4981 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5041 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5101 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5161 gagataccta cagcgtgagc tatgaga
//
