LOCUS       sequence-469258-        6739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  over-express E. coli dnaN (Pol III beta sliding clamp subunit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6739)
  AUTHORS   O'Donnell Lab
  TITLE     O'Donnell Lab Batch 1
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6739)
  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            "unpublished"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 dnaN-pET(11a)
FEATURES             Location/Qualifiers
     source          1..6739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             784..975
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(969..991)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1077..1216
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1132..1151
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1231..1248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1402..1990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1482..1501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2161..2952,2953..3021))
                     /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     2784..2803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3022..3126)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3194..3212
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(3239..3244,3245..3261,3262..3267)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(3442..3489)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3485..3503
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             4662..4684
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4699..4723
                     /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(4723..4742)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4724..4742)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4826..4845)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5051..5128
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5129..6211
                     /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(5148..5167)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
       61 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
      121 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
      181 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
      241 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
      301 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
      361 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
      421 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
      481 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc
      541 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
      601 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta
      661 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc
      721 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca
      781 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca
      841 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa
      901 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac
      961 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     1021 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     1081 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     1141 agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat gcgtaaggag
     1201 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1801 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     1861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1921 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1981 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2041 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2101 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2161 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2221 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2281 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2341 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2401 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2461 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2521 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2581 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2641 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2701 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2761 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     2821 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2881 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2941 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3001 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3061 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3121 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3181 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tctcatgttt
     3241 gacagcttat catcgataag ctttaatgcg gtagtttatc acagttaaat tgctaacgca
     3301 gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc
     3361 tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcc
     3421 ggatatagtt cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     3481 ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt
     3541 agcagccgga tccttacagt ctcattggca tgacaacata agccgcgctc tggctggccg
     3601 catcttcaat ctgcacgctg gaaaccgaat cggtcagcat catgcggacg ttttcgcatt
     3661 tcagcgcgtt cagaacatcc agcacataac tgacgttgaa gccgatttcc atctccgcac
     3721 cgctataggt aacgtcgagg atctcttccg cttcttcctg ttccgggttg ttggcggtga
     3781 ttttcagctg gttttcgctg acataaagac gtacgccgcg gaatttctcg ttagagagaa
     3841 tcgccgcgcg agcaaacgcc tgcttgagca gatcgcagcc agcttccaga tgtttgtccg
     3901 ggttcttcgg cagaacgcgg cgataatccg ggaagcgacc atccaccagt ttggaggtga
     3961 agataaagtc gccaacgtgg gcgcgaatgt tgttgctgcc aatctgtacg cgcagcggat
     4021 tgtcgccgcc gtcgagcata cgcatcagtt caatcacgcc tttacgcggt acgatcaccg
     4081 aatggcttgg caaagattga ccaattggca ttgaacagac cgccagacgg tggccgtcgg
     4141 ttgccacggt gcgcagttct tcaccttcgg tttcaaacag cataccattt aagtaatagc
     4201 gaacgtcctg atgcgccata gaaaactggg tcgcttcaat cagacgcttc atcgttgcct
     4261 gcggcagggt aaattcgact tcactctgcc agtcatcgag gttcgggaaa tccgccgctg
     4321 gcagggtaga cagcgaaaaa cggctacgcc cggagcgtac cagcatccgt tcaccttcca
     4381 gctgcacggc aatttccgcg ccttcaggca gaccacggca gatatcaaag aatttgcgcg
     4441 ccggaacggt cgtcgctcct ggctcgtgtg gctgaaccag cgcaacacgt gccaccattt
     4501 ccatctcgag atcagtaccg gtcagcgaca acgtaccgtc agcaacctgt aacagcagat
     4561 taccgagaat cggtagcgta ggacgaccac ctaacggacc gctcacctgt tgtagcggtt
     4621 ttaataaatg ctcacgttct acggtaaatt tcatatgtat atctccttct taaagttaaa
     4681 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
     4741 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
     4801 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
     4861 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
     4921 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
     4981 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     5041 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     5101 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     5161 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     5221 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     5281 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     5341 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     5401 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     5461 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     5521 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     5581 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     5641 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     5701 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     5761 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     5821 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     5881 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     5941 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     6001 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     6061 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     6121 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6181 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     6241 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     6301 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     6361 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     6421 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     6481 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     6541 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     6601 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     6661 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     6721 ttaccagcct aacttcgat
//
