LOCUS       sequence-267962-        6635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SSB-C-term-GFP fusion expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6635)
  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 6635)
  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: pKD072
FEATURES             Location/Qualifiers
     source          1..6635
                     /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             1367..1384
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1437..1455)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1451..1498
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1535..1990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1622..1641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1832..1853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2016..2120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2121..2189,2190..2981)
                     /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(2339..2358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3152..3740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3641..3660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3894..3911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3926..4065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3991..4010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4151..4173
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4167..4358)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5167..6249)
                     /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     6211..6230
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6250..6327)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6533..6552
                     /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 tcagcggcgg
      541 cgcgcagtct cgcccgcagc agtccgctcc ggcagcgccg tctaacgagc cgccgatgga
      601 ctttgatgat gacattccgt tcggtagcat cagtaaaaga gaagaactgt tcaccggtgt
      661 tgttccgatc ctggttgaac tggatggtga tgttaacggc cacaaattct ctgttcgtgg
      721 tgaaggtgaa ggtgatgcaa ccaacggtaa actgaccctg aaattcatct gcactaccgg
      781 taaactgccg gttccatggc cgactctggt gactaccctg acctatggtg ttcagtgttt
      841 ttctcgttac ccggatcaca tgaagcagca tgatttcttc aaatctgcaa tgccggaagg
      901 ttatgtacag gagcgcacca tttctttcaa agacgatggc acctacaaaa cccgtgcaga
      961 ggttaaattt gaaggtgata ctctggtgaa ccgtattgaa ctgaaaggca ttgatttcaa
     1021 agaggacggc aacatcctgg gccacaaact ggaatataac ttcaactccc ataacgttta
     1081 catcaccgca gacaaacaga agaacggtat caaagctaac ttcaaaattc gccataacgt
     1141 tgaagacggt agcgtacagc tggcggacca ctaccagcag aacactccga tcggtgatgg
     1201 tccggttctg ctgccggata accactacct gtccacccag tctaaactgt ccaaagaccc
     1261 gaacgaaaag cgcgaccaca tggtgctgct ggagttcgtt actgcagcag gtatcacgca
     1321 cggcatggat gagctctaca aatgaaagct tgcggccgca ctcgagcacc accaccacca
     1381 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1441 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1501 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1681 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1741 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1801 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1861 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1921 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1981 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2041 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2101 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2161 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2221 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2281 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2341 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2401 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2461 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2521 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2581 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2641 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2701 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2761 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2821 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2881 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2941 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3001 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3061 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3121 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3181 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3241 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3301 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3361 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3421 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3481 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3541 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3601 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3661 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3721 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3781 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3841 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3901 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3961 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4021 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     4081 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4141 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4201 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     4261 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     4321 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     4381 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     4441 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4501 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4561 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4621 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4681 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4741 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4801 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
     4861 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4921 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4981 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     5041 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     5101 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     5161 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5221 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5281 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5341 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5401 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5461 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5521 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5581 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5641 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5701 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5761 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5821 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5881 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5941 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     6001 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     6061 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     6121 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     6181 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     6241 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     6301 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     6361 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     6421 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6481 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6541 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6601 cggcgtagag gatcgagatc tcgatcccgc gaaat
//
