LOCUS       sequence-469263-        5516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  over-express E. coli holD (Pol III psi subunit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5516)
  AUTHORS   O'Donnell Lab
  TITLE     O'Donnell Lab Batch 1
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5516)
  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            "unpublished"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 holD-pET(3c)
FEATURES             Location/Qualifiers
     source          1..5516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..46)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    132..271
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     187..206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(286..303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(457..1045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(537..556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1216..2007,2008..2076))
                     /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     1839..1858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2077..2181)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2249..2267
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2294..2299,2300..2316,2317..2322)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2688..2735)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2731..2749
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             3722..3744
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3775..3794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3776..3794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3878..3897)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             join(5355..5516,1..30)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
       61 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      121 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      181 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
      241 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
      301 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      361 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      421 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      481 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      541 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      601 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      661 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      721 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      781 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      841 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
      901 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      961 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1021 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1081 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1141 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1201 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1261 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1321 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1381 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1441 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1501 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     1561 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1621 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1681 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1741 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1801 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     1861 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1921 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1981 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2041 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2101 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2161 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2221 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     2281 agaattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt
     2341 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
     2401 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
     2461 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc
     2521 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg
     2581 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
     2641 cgaccacacc cgtcctgtgg atatccggat atagttcctc ctttcagcaa aaaacccctc
     2701 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
     2761 aactcagctt cctttcgggc tttgttagca gccggatccc tgcaggccgg tgaatgagtc
     2821 aaaggtaaac agcgtttcat cggcatcaaa gaaaatccag tcccacttca ttattccacc
     2881 ttgtattaca tactgattgg caacgccatg atgatggcgt cttcgcgacc gtccgtggtg
     2941 gggtagtaat tgcggcgaat cgtcgcctcg ttaaagccta aactttcgta cagggcaatg
     3001 gcggcagcgt ttgaagcacg gacttccagc catagtgtcg ccacgccgcg tttttccagt
     3061 tcgtcgatca gatgttccag cagcgccctt cccaatccct gacgctgata gtcaggatcg
     3121 accgcaatat tgaacaatgt agcttcatcc agcaccactt gcgtaatcgc aaacgccgcc
     3181 attttgccgt tttgcgttaa ctgaaagttg agataacgct cgccctggtt gctggcaaac
     3241 gttttttcac tccacggaaa ggcgtgggcg cgttgttcaa tgtggtaagc cgccggtaaa
     3301 tcagtcgttt cgagggaaga aatcgtgttc atatgtgcaa atttgttgcc ataacgcggc
     3361 gcgtgccgtt gggtttgccc gtaaatcggt gagcgccggt gatgccacct gagcgccttc
     3421 cagtgatagc ggttcgtcag tacccaaccg ccaactgttg cagtgactgc cttgcggcag
     3481 catcgcgatt ttttctggcg tcagttgcag cacctggtcg gggctgacgg ttaatgcgcg
     3541 cagaacatcg ctcactaaag gatcagtcag ggcgggaaga tcgtttgcca ccatcaccag
     3601 acggacgtgt gccgggatcg caatggcaat ctccccctgc aacgcgccag ggcgacgcag
     3661 cgaccactgg gtaatgccca gttgctgtaa ctgccagtct cgtcgggatg tcatatgtat
     3721 atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg gtctccctat
     3781 agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg acgcatcgtg
     3841 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     3901 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     3961 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     4021 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     4081 aagggagagc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     4141 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     4201 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     4261 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     4321 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     4381 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc
     4441 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc
     4501 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc
     4561 ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca
     4621 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg
     4681 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta
     4741 acggattcac cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc
     4801 aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc
     4861 gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga
     4921 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     4981 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     5041 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     5101 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     5161 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     5221 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     5281 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     5341 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     5401 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     5461 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcg
//
