LOCUS       sequence-266428-        5561 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 5561)
  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 5561)
  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..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     206..225
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     309..328
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        309..327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    328..352
                     /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             367..389
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             730..747
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(800..818)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      814..861
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      898..1353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(985..1004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1195..1216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1446..2261)
                     /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(1446..2261)
                     /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     2169..2188
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2253..2272
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2383..2971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2872..2891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3125..3142
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3157..3299
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3225..3244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3385..3407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3401..3592)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4401..5483)
                     /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     5445..5464
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5484..5561)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
       61 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      121 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      181 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      241 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      301 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      361 aataattttg tttaacttta agaaggagat ataccatggc gtatagcatt accctgcgct
      421 cgccggatgg cgctgaagaa gtggtacagt gcgaggaaga tcagtatatt ttagaagcgg
      481 cggaggatgc tgggctggat atgccatcaa gttgccgtgc gggcgcttgc tccgcttgtc
      541 tcggcaaagt gttagagggg tcagttaaca atgaggaaca gtctttttta gacgacgatc
      601 agctggaaga aggatggtca ctgctttgcg tggcaatgcc acagtcggac tgtgttatcc
      661 tgacggagca agaagacaac ctggattaag aattcgagct ccgtcgacaa gcttgcggcc
      721 gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
      781 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      841 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
      901 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      961 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1021 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1081 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1141 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1201 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1261 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1321 cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc
     1381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     1441 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1501 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1561 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1621 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1681 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1741 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1801 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1861 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1921 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1981 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2041 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2101 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2161 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2221 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     2281 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     2341 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2401 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2461 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2521 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2581 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2641 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2701 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2761 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2821 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2881 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2941 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3001 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3061 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3121 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3181 gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat
     3241 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3301 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3361 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3421 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     3481 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     3541 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     3601 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     3661 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     3721 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     3781 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     3841 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     3901 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     3961 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     4021 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     4081 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     4141 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     4201 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     4261 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     4321 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     4381 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     4501 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     4561 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4621 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4681 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4741 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4801 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4861 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4921 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4981 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5041 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5101 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5161 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5221 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5281 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     5341 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     5401 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     5461 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     5521 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt c
//
