LOCUS       sequence-267963-        6660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SSB-GFP IDL fusion expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6660)
  AUTHORS   Dubiel K, Henry C, Spenkelink LM, Kozlov AG, Wood EA, Jergic S, 
            Dixon NE, van Oijen AM, Cox MM, Lohman TM, Sandler SJ, Keck JL
  TITLE     Development of a single-stranded DNA-binding protein fluorescent 
            fusion toolbox.
  JOURNAL   Nucleic Acids Res. 2020 Jun 19;48(11):6053-6067. doi: 
            10.1093/nar/gkaa320.
  PUBMED    32374866
REFERENCE   2  (bases 1 to 6660)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2020-06-19- 19"; volume: "48"; issue: 
            "11"; pages: "6053-6067"
COMMENT     Sequence Label: pKD068
FEATURES             Location/Qualifiers
     source          1..6660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1392..1409
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1462..1480)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1476..1523
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1560..2015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1647..1666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1857..1878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2041..2145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2146..2214,2215..3006)
                     /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(2364..2383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3177..3765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3666..3685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3919..3936
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3951..4090
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4016..4035)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4176..4198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4192..4383)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5192..6274)
                     /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     6236..6255
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6275..6352)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6558..6577
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggccagcaga ggcgtaaaca aggttattct
      121 cgttggtaat ctgggtcagg acccggaagt acgctacatg ccaaatggtg gcgcagttgc
      181 caacattacg ctggctactt ccgaatcctg gcgtgataaa gcgaccggcg agatgaaaga
      241 acagactgaa tggcaccgcg ttgtgctgtt cggcaaactg gcagaagtgg cgagcgaata
      301 tctgcgtaaa ggttctcagg tttatatcga aggtcagctg cgtacccgta aatggaccga
      361 tcaatccggt caggatcgct acaccacaga agtcgtggtg aacgttggcg gcaccatgca
      421 gatgctgggt ggtcgtcagg gtggtggcgc tccggcaggt ggcaatatcg gtggtggtca
      481 gccgcagggc ggttggggtc agcctcagca gccgcagggt ggcaatcagt tcggtagcat
      541 cagtaaaaga gaagaactgt tcaccggtgt tgttccgatc ctggttgaac tggatggtga
      601 tgttaacggc cacaaattct ctgttcgtgg tgaaggtgaa ggtgatgcaa ccaacggtaa
      661 actgaccctg aaattcatct gcactaccgg taaactgccg gttccatggc cgactctggt
      721 gactaccctg acctatggtg ttcagtgttt ttctcgttac ccggatcaca tgaagcagca
      781 tgatttcttc aaatctgcaa tgccggaagg ttatgtacag gagcgcacca tttctttcaa
      841 agacgatggc acctacaaaa cccgtgcaga ggttaaattt gaaggtgata ctctggtgaa
      901 ccgtattgaa ctgaaaggca ttgatttcaa agaggacggc aacatcctgg gccacaaact
      961 ggaatataac ttcaactccc ataacgttta catcaccgca gacaaacaga agaacggtat
     1021 caaagctaac ttcaaaattc gccataacgt tgaagacggt agcgtacagc tggcggacca
     1081 ctaccagcag aacactccga tcggtgatgg tccggttctg ctgccggata accactacct
     1141 gtccacccag tctaaactgt ccaaagaccc gaacgaaaag cgcgaccaca tggtgctgct
     1201 ggagttcgtt actgcagcag gtatcacgca cggcatggat gagctctaca aaagcggcgg
     1261 cgcgcagtct cgcccgcagc agtccgctcc ggcagcgccg tctaacgagc cgccgatgga
     1321 ctttgatgat gacattccgt tctaaggatc cgaattcgag ctccgtcgac aagcttgcgg
     1381 ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     1441 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1501 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     1561 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1621 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1681 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1741 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1801 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1861 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1921 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1981 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     2041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2521 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     2701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3001 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3301 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3841 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3901 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3961 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     4021 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4081 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4141 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4201 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4261 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4321 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4381 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     4441 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4501 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4561 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     4621 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     4681 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     4741 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     4801 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     4861 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     4921 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     4981 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     5041 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     5101 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     5161 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5221 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5281 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     5341 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     5401 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     5461 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     5521 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     5581 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     5641 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     5701 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     5761 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     5821 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     5881 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     5941 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     6001 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     6061 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     6121 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     6181 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     6241 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     6301 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     6361 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     6421 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     6481 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     6541 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     6601 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
