LOCUS       sequence-324164-        6043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. elongatus Form IB rubisco and S. elongatus Prk.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6043)
  AUTHORS   Flamholz AI, Dugan E, Blikstad C, Gleizer S, Ben-Nissan R, Amram S, 
            Antonovsky N, Ravishankar S, Noor E, Bar-Even A, Milo R, Savage D
  TITLE     Functional reconstitution of a bacterial CO2 concentrating mechanism
            in E. coli.
  JOURNAL   Elife. 2020 Oct 21;9. pii: 59882. doi: 10.7554/eLife.59882.
  PUBMED    33084575
REFERENCE   2  (bases 1 to 6043)
  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: "Elife."; 
            date: "2020-10-21"; volume: "9. pii: 59882. doi"; pages: " 
            10.7554/eLife.59882"
COMMENT     Sequence Label: pFE-Selo-FI-prk
FEATURES             Location/Qualifiers
     source          1..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(162..750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(242..261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(838..932)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(963..1757)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1074..1093)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1684..1703
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1936..1954
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1975..1993,1994..1999,2000..2016,2017..2022,
                     2023..2048)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1975..1993
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2000..2018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2058..2069
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3975..3992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5042..5128
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5301..5924)
                     /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     5832..5851
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(5925..6029)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgctaga cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
       61 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      601 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgactag tgcttggatt
      841 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
      901 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc
      961 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     1021 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     1081 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     1141 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     1201 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     1261 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     1321 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     1381 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     1441 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     1501 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     1561 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     1621 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     1681 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     1741 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     1801 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     1861 accttaccag agggcgcccc agctggcaat tccgacgtct aagaaaccat tattatcatg
     1921 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagtcccta
     1981 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacat cagcaggacg
     2041 cactgaccga attcattaaa gaggagaaag gtaccatggc caagacgcaa tctgccgcag
     2101 gctataaggc cggggtgaag gactacaaac tcacctatta cacccccgat tacaccccca
     2161 aagacactga cctgctggcg gctttccgct tcagccctca gccgggtgtc cctgctgacg
     2221 aagctggtgc ggcgatcgcg gctgaatctt cgaccggtac ctggaccacc gtgtggaccg
     2281 acttgctgac cgacatggat cggtacaaag gcaagtgcta ccacatcgag ccggtgcaag
     2341 gcgaagagaa ctcctacttt gcgttcatcg cttacccgct cgacctgttt gaagaagggt
     2401 cggtcaccaa catcctgacc tcgatcgtcg gtaacgtgtt tggcttcaaa gctatccgtt
     2461 cgctgcgtct ggaagacatc cgcttccccg tcgccttggt caaaaccttc caaggtcctc
     2521 cccacggtat ccaagtcgag cgcgacctgc tgaacaagta cggccgtccg atgctgggtt
     2581 gcacgatcaa accaaaactc ggtctgtcgg cgaaaaacta cggtcgtgcc gtctacgaat
     2641 gtctgcgcgg cggtctggac ttcaccaaag acgacgaaaa catcaactcg cagccgttcc
     2701 aacgctggcg cgatcgcttc ctgtttgtgg ctgatgcaat ccacaaatcg caagcagaaa
     2761 ccggtgaaat caaaggtcac tacctgaacg tgaccgcgcc gacctgcgaa gaaatgatga
     2821 aacgggctga gttcgctaaa gaactcggca tgccgatcat catgcatgac ttcttgacgg
     2881 ctggtttcac cgccaacacc accttggcaa aatggtgccg cgacaacggc gtcctgctgc
     2941 acatccaccg tgcaatgcac gcggtgatcg accgtcagcg taaccacggg attcacttcc
     3001 gtgtcttggc caagtgtttg cgtctgtccg gtggtgacca cctccactcc ggcaccgtcg
     3061 tcggcaaact ggaaggcgac aaagcttcga ccttgggctt tgttgacttg atgcgcgaag
     3121 accacatcga agctgaccgc agccgtgggg tcttcttcac ccaagattgg gcgtcgatgc
     3181 cgggcgtgct gccggttgct tccggtggta tccacgtgtg gcacatgccc gcactggtgg
     3241 aaatcttcgg tgatgactcc gttctccagt tcggtggcgg caccttgggt cacccctggg
     3301 gtaatgctcc tggtgcaacc gcgaaccgtg ttgccttgga agcttgcgtc caagctcgga
     3361 acgaaggtcg cgacctctac cgtgaaggcg gcgacatcct tcgtgaagct ggcaagtggt
     3421 cgcctgaact ggctgctgcc ctcgacctct ggaaagagat caagttcgaa ttcgaaacga
     3481 tggacaagct ctaaggagcc tctgactatc gctgggggag tgagcgttgc tgcgtaaagc
     3541 tttctcccca gcctttcgac ttaacctttc aggatttctg aatcatgagc atgaaaactc
     3601 tgcccaaaga gcgtcgtttc gagactttct cgtacctgcc tcccctcagc gatcgccaaa
     3661 tcgctgcaca aatcgagtac atgatcgagc aaggcttcca ccccttgatc gagttcaacg
     3721 agcactcgaa tccggaagag ttctactgga cgatgtggaa gctccccctg tttgactgca
     3781 agagccctca gcaagtcctc gatgaagtgc gtgagtgccg cagcgaatac ggtgattgct
     3841 acatccgtgt cgctggcttc gacaacatca agcagtgcca aaccgtgagc ttcatcgttc
     3901 atcgtcccgg ccgctactaa taaatctaat agaaataatt ttgtttaact ttataagcag
     3961 gaccggcggc gatgcatcat caccatcacc acagcaagcc agatcgtgtt gttttgatcg
     4021 gcgttgccgg tgactccggt tgcggcaaat caaccttcct aaatcgcctt gccgacttgt
     4081 ttggtacgga attgatgacg gtcatctgct tggatgacta tcacagtctc gatcgcaagg
     4141 gccggaagga agcaggcgta acggctttgg atccccgcgc caacaacttt gacttgatgt
     4201 atgaacaggt caaggcgttg aagaacggcg aaacgatcat gaagccgatc tacaaccatg
     4261 aaaccggctt gatcgatccg cccgaaaaaa tcgaacccaa tcgcatcatt gtgatcgagg
     4321 gtctgcatcc gctttacgac gagcgcgtgc gtgaactgct cgatttcagc gtttacctcg
     4381 acatcgatga cgaagtcaaa atcgcttgga agatccaacg cgatatggca gaacgcggcc
     4441 actcctacga agatgtcctc gcctcgatcg aagcgcgccg ccctgacttc aaggcctaca
     4501 ttgagcccca gcgtggccat gcggacatcg tcatccgcgt catgccgacc cagctaatcc
     4561 ccaatgacac cgagcgcaag gtgctgcggg tgcagttgat ccaacgggaa ggccgcgatg
     4621 gttttgagcc ggcttacctg ttcgacgaag gttcgaccat ccagtggacg ccctgcggtc
     4681 gtaagctgac ctgctcctat ccgggcattc gcttagccta cggccctgac acctactacg
     4741 gtcacgaagt ctcagtgctt gaggtcgacg gtcagttcga gaacctcgaa gagatgatct
     4801 acgtcgaggg ccacctcagc aagaccgaca cgcagtacta cggtgagttg acccacctgc
     4861 tgctacagca caaagattac ccgggttcga acaacggcac gggtctgttc caagtgctga
     4921 ccggcctgaa aatgcgggcg gcctatgagc gtttgacctc ccaagcagca cccgtcgccg
     4981 ctagtgtcta gatggaattc ctgcagcccg ggggatccca tggtacgcgt gctagaggca
     5041 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     5101 ggtgaacgct ctcctgagta ggacaaatcc gccgccgcta gcagttcgta gacggaaaca
     5161 aacgcagaat ccaagcaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5221 ttcctttttc aatattattg aagcatttat cagggttatg tccactgggt tcgtgccttc
     5281 atccggatat ccaggaactg ttaagaccca ctttcacatt taagttgttt ttctaatccg
     5341 catatgatca attcaaggcc gaataagaag gctggctctg caccttggtg atcaaataat
     5401 tcgatagctt gtcgtaataa tggcggcata ctatcagtag taggtgtttc cctttcttct
     5461 ttagcgactt gatgctcttg atcttccaat acgcaaccta aagtaaaatg ccccacagcg
     5521 ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga
     5581 ttttcgagag tttcatactg tttttctgta ggccgtgtac ctaaatgtac ttttgctcca
     5641 tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt tattacgtaa aaaatcttgc
     5701 cagctttccc cttctaaagg gcaaaagtga gtatggtgcc tatctaacat ctcaatggct
     5761 aaggcgtcga gcaaagcccg cttatttttt acatgccaat acaatgtagg ctgctctaca
     5821 cctagcttct gggcgagttt acgggttgtt aaaccttcga ttccgacctc attaagcagc
     5881 tctaatgcgc tgttaatcac tttactttta tctaatctag acatactctt cctttttcaa
     5941 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6001 tagaaaaata aacaaatagg ggttccgcgt ttccctcgac aat
//
