LOCUS       sequence-273510-        3912 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 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: pFd030.R2.RFP
FEATURES             Location/Qualifiers
     source          1..3912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..19,20..25,26..42,43..48,49..74)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             433..1107
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      1137..1208
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1224..1251
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1303..1337
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1372..1998
                     /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(1448..1467)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2004..2090
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2255..2843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2335..2354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2931..3025)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3046..3705)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3620..3639
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3874..3892
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc
       61 aggacgcact gaccgattac cggcaacagg agacacaaaa tgacgagctt tcaaattacc
      121 tttattaacg agttatccgg gttgaaggag attatcgata ttccagatga caagtacatt
      181 ctcgatgcag cttgtgagca gaatattgaa ctcccggcgt catgtcgtga agggacttgt
      241 tcaacctgtg tcgccaagtt ggagaagggg aatgtcgacc aacaggacca gtctttcctt
      301 gacgatgatc aaatcgaaaa gggctacatt ttgatttgca aggcctaccc tctgagtgat
      361 tgcattatcc gcacacatgc ggaagatcaa ctttacgggg gctctggagg ttctggaggt
      421 tctggtgggt ccgcgagtag cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt
      481 atggaaggtt ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg
      541 tacgaaggta cccagaccgc taaactgaaa gttaccaaag gtggtccgct gccgttcgct
      601 tgggacatcc tgtccccgca gttccagtac ggttccaaag cttacgttaa acacccggct
      661 gacatcccgg actacctgaa actgtccttc ccggaaggtt tcaaatggga acgtgttatg
      721 aacttcgaag acggtggtgt tgttaccgtt acccaggact cctccctgca agacggtgag
      781 ttcatctaca aagttaaact gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag
      841 aaaaaaacca tgggttggga agcttccacc gaacgtatgt acccggaaga cggtgctctg
      901 aaaggtgaaa tcaaaatgcg tctgaaactg aaagacggtg gtcactacga cgctgaagtt
      961 aaaaccacct acatggctaa aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc
     1021 aaactggaca tcacctccca caacgaagac tacaccatcg ttgaacagta cgaacgtgct
     1081 gaaggtcgtc actccaccgg tgcttaagaa ttctgataat aatctagacc aggcatcaaa
     1141 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1201 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttataggt
     1261 acccgtggat cctctagagt cgacctgcag gcatgcaagc tttttacggc tagctcagtc
     1321 ctaggtatag tgctagccca gccagagaaa cactctttaa cagggggtag tatgatgtct
     1381 cgtttagata aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc
     1441 gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat
     1501 tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg
     1561 caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac
     1621 gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt
     1681 acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa
     1741 caaggttttt cactagagaa tgcattatat gcactcagcg cagtggggca ttttacttta
     1801 ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact
     1861 actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca
     1921 gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa acaacttaaa
     1981 tgtgaaagtg ggtcttaagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2041 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
     2101 agacctaggg cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2161 tatccacaga atcaggggat aacgcatgca agaacatgtg agcaaaaggc cagcaaaagg
     2221 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2281 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2341 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2401 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2461 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2521 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2581 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2641 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     2701 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2761 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2821 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2881 agtggaacga aaactcacgt taagggattt tggtcatgac tagtgcttgg attctcacca
     2941 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     3001 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     3061 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     3121 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     3181 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     3241 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     3301 agggaaatag gccaggtttt caccataaca cgccacatct tgcgaatata tgtgtagaaa
     3361 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     3421 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     3481 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     3541 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     3601 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     3661 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     3721 tcctgaaaat ctcgataact caaaaaatac gaccggtagt gatcttattt cattatggtg
     3781 aaagttggaa cctcttacgg gccggccaac gtctcatttt cgccagatat cgacgtctaa
     3841 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3901 cttcacctcg ag
//
