LOCUS       sequence-297212-        3714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA carrying a sfGFP gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3714)
  AUTHORS   Desmarais JJ, Flamholz AI, Blikstad C, Dugan EJ, Laughlin TG, 
            Oltrogge LM, Chen AW, Wetmore K, Diamond S, Wang JY, Savage DF
  TITLE     DABs are inorganic carbon pumps found throughout prokaryotic phyla.
  JOURNAL   Nat Microbiol. 2019 Aug 12. pii: 10.1038/s41564-019-0520-8. doi: 
            10.1038/s41564-019-0520-8.
  PUBMED    31406332
REFERENCE   2  (bases 1 to 3714)
  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: "Nat 
            Microbiol. 2019 Aug 12. pii: 10.1038/s41564-019-0520-8. doi: 
            10.1038/s41564-019-0520-8."
COMMENT     Sequence Label: pFA-GFP
FEATURES             Location/Qualifiers
     source          1..3714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(113..130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(247..792)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(327..346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(906..1000)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1021..1680)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1595..1614
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1681..1783)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1849..1867
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1888..1906,1907..1912,1913..1929,1930..1935,
                     1936..1961)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1888..1906
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1913..1931
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1971..1982
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1989..1991,1992..1994,1995..1997,1998..2708)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="MSVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGI
                     KANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     primer_bind     complement(2037..2058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2645..2666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2715..2732
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2794..2880
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3053..3676)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     3584..3603
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(join(3677..3714,1..67))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
       61 ttccgcgttt ccctcgacaa ttcgcgctag acctagggat atattccgct tcctcgctca
      121 ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg
      181 gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa
      241 gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca
      301 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct
      361 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
      421 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
      481 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
      541 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
      601 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
      661 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
      721 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
      781 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
      841 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gttactagtg
      901 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
      961 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     1021 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1081 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1141 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     1201 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     1261 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     1321 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1381 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1441 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg
     1501 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     1561 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     1621 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     1681 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     1741 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     1801 gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     1861 acgaggccct ttcgtcttca cctcgagtcc ctatcagtga tagagattga catccctatc
     1921 agtgatagag atactgagca catcagcagg acgcactgac cgaattcatt aaagaggaga
     1981 aaggtaccat gtccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2041 tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag ggcgagggcg
     2101 atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2161 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2221 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2281 gcaccatctc cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg
     2341 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2401 tcctggggca caagctggag tacaacttca acagccacaa cgtctatatc acggccgaca
     2461 agcagaagaa cggcatcaag gcgaacttca agatccgcca caacgtcgag gacggcagcg
     2521 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2581 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     2641 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2701 tgtacaagag cggtcaccat caccatcacc attaaaatgt aactgtattc agcgatgacg
     2761 aaattcttag ctattgtaat actctagagg catcaaataa aacgaaaggc tcagtcgaaa
     2821 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     2881 ccgccgccgc tagcagttcg tagacggaaa caaacgcaga atccaagcaa aagggaataa
     2941 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3001 atcagggtta tgtccactgg gttcgtgcct tcatccggat atccaggaac tgttaagacc
     3061 cactttcaca tttaagttgt ttttctaatc cgcatatgat caattcaagg ccgaataaga
     3121 aggctggctc tgcaccttgg tgatcaaata attcgatagc ttgtcgtaat aatggcggca
     3181 tactatcagt agtaggtgtt tccctttctt ctttagcgac ttgatgctct tgatcttcca
     3241 atacgcaacc taaagtaaaa tgccccacag cgctgagtgc atataatgca ttctctagtg
     3301 aaaaaccttg ttggcataaa aaggctaatt gattttcgag agtttcatac tgtttttctg
     3361 taggccgtgt acctaaatgt acttttgctc catcgcgatg acttagtaaa gcacatctaa
     3421 aacttttagc gttattacgt aaaaaatctt gccagctttc cccttctaaa gggcaaaagt
     3481 gagtatggtg cctatctaac atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt
     3541 ttacatgcca atacaatgta ggctgctcta cacctagctt ctgggcgagt ttacgggttg
     3601 ttaaaccttc gattccgacc tcattaagca gctctaatgc gctgttaatc actttacttt
     3661 tatctaatct agacatactc ttcctttttc aatattattg aagcatttat cagg
//
