LOCUS       sequence-273513-        3204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Prochlorococcus marinus NATL2A Fd 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3204)
  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 3204)
  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
FEATURES             Location/Qualifiers
     source          1..3204
                     /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"
     terminator      429..500
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      516..543
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        595..629
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             664..1290
                     /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(740..759)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1296..1382
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1547..2135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1627..1646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2223..2317)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2338..2997)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2912..2931
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3166..3184
                     /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 ctttactaag aattctgata ataatctaga
      421 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      481 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
      541 gcgtttatag gtacccgtgg atcctctaga gtcgacctgc aggcatgcaa gctttttacg
      601 gctagctcag tcctaggtat agtgctagcc cagccagaga aacactcttt aacagggggt
      661 agtatgatgt ctcgtttaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat
      721 gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag
      781 cctacattgt attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag
      841 atgttagata ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt
      901 ttacgtaata acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa
      961 gtacatttag gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc
     1021 tttttatgcc aacaaggttt ttcactagag aatgcattat atgcactcag cgcagtgggg
     1081 cattttactt taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg
     1141 gaaacaccta ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat
     1201 caccaaggtg cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa
     1261 aaacaactta aatgtgaaag tgggtcttaa ggcatcaaat aaaacgaaag gctcagtcga
     1321 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1381 atccgccgcc ctagacctag ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1441 cggtaatacg gttatccaca gaatcagggg ataacgcatg caagaacatg tgagcaaaag
     1501 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1561 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1621 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1681 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1741 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1801 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1861 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1921 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1981 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2041 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2101 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2161 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg actagtgctt
     2221 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     2281 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
     2341 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2401 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2461 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2521 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2581 aataaaccct ttagggaaat aggccaggtt ttcaccataa cacgccacat cttgcgaata
     2641 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2701 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2761 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
     2821 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     2881 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     2941 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     3001 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgaccggta gtgatcttat
     3061 ttcattatgg tgaaagttgg aacctcttac gggccggcca acgtctcatt ttcgccagat
     3121 atcgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3181 aggccctttc gtcttcacct cgag
//
