LOCUS       sequence-266415-        3912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Prochlorococcus phage P-SSM2 phage Fd fused to mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3912)
  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 3912)
  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: pFd022.R2.RFP
FEATURES             Location/Qualifiers
     source          1..3912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(194..288)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(309..968)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     883..902
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(969..1071)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1137..1155
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1176..1194,1195..1200,1201..1217,1218..1223,
                     1224..1249)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1176..1194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1201..1219
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1609..2283
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2313..2384
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2400..2427
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2479..2513
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2548..3174
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2624..2643)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3180..3266
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(3430..3912,1..106))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3510..3529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
       61 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      121 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      181 gactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
      241 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
      301 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
      361 ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc
      421 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
      481 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
      541 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
      601 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
      661 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
      721 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
      781 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
      841 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
      901 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
      961 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     1021 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
     1081 tttcgccaga tatcgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     1141 ggcgtatcac gaggcccttt cgtcttcacc tcgagtccct atcagtgata gagattgaca
     1201 tccctatcag tgatagagat actgagcaca tcagcaggac gcactgaccg tataaacggg
     1261 gaatatagaa aagagggttc tatggcgtat agcattaccc tgcgctcgcc ggatggcgct
     1321 gaagaagtgg tacagtgcga ggaagatcag tatattttag aagcggcgga ggatgctggg
     1381 ctggatatgc catcaagttg ccgtgcgggc gcttgctccg cttgtctcgg caaagtgtta
     1441 gaggggtcag ttaacaatga ggaacagtct tttttagacg acgatcagct ggaagaagga
     1501 tggtcactgc tttgcgtggc aatgccacag tcggactgtg ttatcctgac ggagcaagaa
     1561 gacaacctgg atgggggctc tggaggttct ggaggttctg gtgggtccgc gagtagcgaa
     1621 gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac
     1681 gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa
     1741 ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc
     1801 cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg
     1861 tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt
     1921 accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt
     1981 ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct
     2041 tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg
     2101 aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa
     2161 ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac
     2221 gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct
     2281 taagaattct gataataatc tagaccaggc atcaaataaa acgaaaggct cagtcgaaag
     2341 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     2401 gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc tagagtcgac
     2461 ctgcaggcat gcaagctttt tacggctagc tcagtcctag gtatagtgct agcccagcca
     2521 gagaaacact ctttaacagg gggtagtatg atgtctcgtt tagataaaag taaagtgatt
     2581 aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc
     2641 gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct
     2701 ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta
     2761 gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta
     2821 ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat
     2881 gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca
     2941 ttatatgcac tcagcgcagt ggggcatttt actttaggtt gcgtattgga agatcaagag
     3001 catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta
     3061 cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt
     3121 gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ttaaggcatc
     3181 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3241 tgaacgctct cctgagtagg acaaatccgc cgccctagac ctaggcgttc ggctgcggcg
     3301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3601 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     3661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3841 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3901 gccagttacc tt
//
