LOCUS       sequence-296241-        5557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inucible expression of Prochlorococcus phage P-SSM2 phage Fd for 
            purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5557)
  AUTHORS   Campbell IJ, Olmos JL Jr, Xu W, Kahanda D, Atkinson JT, Sparks ON, 
            Miller MD, Phillips GN Jr, Bennett GN, Silberg JJ
  TITLE     Prochlorococcus phage ferredoxin: Structural characterization and 
            electron transfer to cyanobacterial sulfite reductases.
  JOURNAL   J Biol Chem. 2020 May 19. pii: RA120.013501. doi: 
            10.1074/jbc.RA120.013501.
  PUBMED    32434930
REFERENCE   2  (bases 1 to 5557)
  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: "J Biol 
            Chem. 2020 May 19. pii: RA120.013501. doi: 
            10.1074/jbc.RA120.013501."
COMMENT     Sequence Label: pJTA022
FEATURES             Location/Qualifiers
     source          1..5557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     151..170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        151..169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    170..194
                     /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             209..231
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             572..589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(642..660)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      656..703
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      740..1195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(827..846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1037..1058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1287..2102)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1287..2102)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2010..2029
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2094..2113
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2224..2812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2713..2732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2966..2983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2998..3137
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3063..3082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3223..3245
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3239..3430)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4239..5321)
                     /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     5283..5302
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5322..5399)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
       61 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
      121 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
      181 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg
      241 gcgtatagca ttaccctgcg ctcgccggat ggcgctgaag aagtggtaca gtgcgaggaa
      301 gatcagtata ttttagaagc ggcggaggat gctgggctgg atatgccatc aagttgccgt
      361 gcgggcgctt gctccgcttg tctcggcaaa gtgttagagg ggtcagttaa caatgaggaa
      421 cagtcttttt tagacgacga tcagctggaa gaaggatggt cactgctttg cgtggcaatg
      481 ccacagtcgg actgtgttat cctgacggag caagaagaca acctggatta agaattcgag
      541 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
      601 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      661 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      721 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      781 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1081 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1141 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     1201 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1261 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     1321 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     1381 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     1441 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     1501 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     1561 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     1621 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     1681 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     1741 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     1801 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     1861 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     1921 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     1981 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2041 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     2101 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
     2161 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2221 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2281 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2341 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2401 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2461 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2521 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2581 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2641 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2701 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2761 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2821 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2881 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2941 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3001 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     3061 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3121 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3181 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3241 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3301 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3361 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3421 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     3481 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     3541 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     3601 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     3661 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     3721 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     3781 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     3841 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     3901 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     3961 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4021 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4081 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4141 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4201 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4321 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     4381 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4441 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     4501 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     4561 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     4621 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     4681 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     4741 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     4801 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     4861 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     4921 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     4981 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5041 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5101 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5161 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     5221 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     5281 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     5341 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     5401 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     5461 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     5521 tcccccggcc acggggcctg ccaccatacc cacgccg
//
