LOCUS       sequence-266422-        3911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Prochlorococcus marinus NATL2A Fd 1 fused to mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3911)
  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 3911)
  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: pFd030.R2.RFP
FEATURES             Location/Qualifiers
     source          1..3911
                     /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             1608..2282
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2312..2383
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2399..2426
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2478..2512
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2547..3173
                     /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(2623..2642)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3179..3265
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(3429..3911,1..106))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3509..3528)
                     /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 attaccggca
     1261 acaggagaca caaaatgacg agctttcaaa ttacctttat taacgagtta tccgggttga
     1321 aggagattat cgatattcca gatgacaagt acattctcga tgcagcttgt gagcagaata
     1381 ttgaactccc ggcgtcatgt cgtgaaggga cttgttcaac ctgtgtcgcc aagttggaga
     1441 aggggaatgt cgaccaacag gaccagtctt tccttgacga tgatcaaatc gaaaagggct
     1501 acattttgat ttgcaaggcc taccctctga gtgattgcat tatccgcaca catgcggaag
     1561 atcaacttta cgggggctct ggaggttctg gaggttctgg tgggtccgcg agtagcgaag
     1621 acgttatcaa agagttcatg cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg
     1681 agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga aggtacccag accgctaaac
     1741 tgaaagttac caaaggtggt ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc
     1801 agtacggttc caaagcttac gttaaacacc cggctgacat cccggactac ctgaaactgt
     1861 ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta
     1921 ccgttaccca ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg
     1981 gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt
     2041 ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga
     2101 aactgaaaga cggtggtcac tacgacgctg aagttaaaac cacctacatg gctaaaaaac
     2161 cggttcagct gccgggtgct tacaaaaccg acatcaaact ggacatcacc tcccacaacg
     2221 aagactacac catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc accggtgctt
     2281 aagaattctg ataataatct agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2341 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     2401 ctcaccttcg ggtgggcctt tctgcgttta taggtacccg tggatcctct agagtcgacc
     2461 tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta gcccagccag
     2521 agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt aaagtgatta
     2581 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     2641 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     2701 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     2761 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     2821 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     2881 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     2941 tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     3001 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     3061 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     3121 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaggcatca
     3181 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3241 gaacgctctc ctgagtagga caaatccgcc gccctagacc taggcgttcg gctgcggcga
     3301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3601 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     3661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3841 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3901 ccagttacct t
//
