LOCUS       sequence-386205-        6179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  wild type E. coli SSB in pET21 expresses in BL21 under T7 control 
            with lactose-amp resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  AUTHORS   Antony E, Weiland E, Yuan Q, Manhart CM, Nguyen B, Kozlov AG, 
            McHenry CS, Lohman TM
  TITLE     Multiple C-terminal tails within a single E. coli SSB homotetramer 
            coordinate DNA replication and repair.
  JOURNAL   J Mol Biol. 2013 Nov 29;425(23):4802-19. doi: 
            10.1016/j.jmb.2013.08.021. Epub 2013 Sep 7.
  PUBMED    24021816
REFERENCE   2  (bases 1 to 6179)
  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"; doi: 
            "10.1016/j.jmb.2013.08.021"; journalName: "J Mol Biol"; date: 
            "2013-11-29- 29"; volume: "425"; issue: "23"; pages: "4802-19"
COMMENT     Sequence Label: SSB-wt
FEATURES             Location/Qualifiers
     source          1..6179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     907..926
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(946..1023)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1229..1248
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1332..1351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1332..1350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1351..1375
                     /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)."
     CDS             2242..2259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2312..2330)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2326..2373
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2410..2865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2497..2516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2707..2728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2891..2995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2996..3064,3065..3856)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3214..3233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4027..4615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4516..4535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4769..4786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4801..4940
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4866..4885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5026..5048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5042..5233)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6042..6179,1..945))
                     /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"
ORIGIN
        1 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
       61 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
      121 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
      181 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
      241 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
      301 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
      361 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
      421 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
      481 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
      541 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
      601 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
      661 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
      721 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
      781 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
      841 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
      901 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
      961 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     1021 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     1081 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     1141 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     1201 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     1261 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     1321 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     1381 agaaacttcg atcttatcca ttcacctttc ccggattaaa cgcttttttg cccggtggca
     1441 tggtgctacc ggcgatcaca aacggttaat tatgacacaa attgacctga atgaatatac
     1501 agtattggaa tgcattaccc ggagtgttgt gtaacaatgt ctggccaggt ttgtttcccg
     1561 gaaccgaggt cacaacatag taaaagcgct attggtaatg gtacaatcgc gcgtttacac
     1621 ttattcagaa cgattttttt tcaggagaca cgaaccatat ggccagcaga ggcgtaaaca
     1681 aggttattct cgttggtaat ctgggtcagg acccggaagt acgctacatg ccaaatggtg
     1741 gcgcagttgc caacattacg ctggctactt ccgaatcctg gcgtgataaa gcgaccggcg
     1801 agatgaaaga acagactgaa tggcaccgcg ttgtgctgtt cggcaaactg gcagaagtgg
     1861 cgagcgaata tctgcgtaaa ggttctcagg tttatatcga aggtcagctg cgtacccgta
     1921 aatggaccga tcaatccggt caggatcgct acaccacaga agtcgtggtg aacgttggcg
     1981 gcaccatgca gatgctgggt ggtcgtcagg gtggtggcgc tccggcaggt ggcaatatcg
     2041 gtggtggtca gccgcagggc ggttggggtc agcctcagca gccgcagggt ggcaatcagt
     2101 tcagcggcgg cgcgcagtct cgcccgcagc agtccgctcc ggcagcgccg tctaacgagc
     2161 cgccgatgga ctttgatgat gacattccgt tctaaggatc cgaattcgag ctccgtcgac
     2221 aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     2281 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2341 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     2401 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2461 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2521 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2581 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2641 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2701 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2761 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2821 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     2881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2941 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3001 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3061 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3121 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3181 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3241 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3301 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3361 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3421 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3481 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     3541 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3601 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3661 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3721 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3781 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3841 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3901 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3961 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4021 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4141 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     4201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4681 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4741 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4801 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     4861 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     4921 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4981 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5041 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5101 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5161 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5221 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     5281 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     5341 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     5401 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     5461 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     5521 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     5581 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     5641 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     5701 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5761 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5821 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5881 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     5941 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     6001 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     6061 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6121 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgatt
//
