LOCUS       sequence-352855-        4792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses heliorhodopsin gene in E.coli cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4792)
  AUTHORS   Chazan A, Rozenberg A, Mannen K, Nagata T, Tahan R, Yaish S, Larom 
            S, Inoue K, Beja O, Pushkarev A
  TITLE     Diverse heliorhodopsins detected via functional metagenomics in 
            freshwater Actinobacteria, Chloroflexi and Archaea.
  JOURNAL   Environ Microbiol. 2022 Jan;24(1):110-121. doi: 
            10.1111/1462-2920.15890. Epub 2022 Jan 4.
  PUBMED    34984789
REFERENCE   2  (bases 1 to 4792)
  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"; doi: 
            "10.1111/1462-2920.15890"; journalName: "Environ Microbiol"; date: 
            "2022-01"; volume: "24"; issue: "1"; pages: "110-121"
COMMENT     Sequence Label: EINA20F1
FEATURES             Location/Qualifiers
     source          1..4792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             312..334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             352..369
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1193..1210)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1363..1449
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1541..1568
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1587..1678
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1679..1747,1748..2539)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1897..1916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2710..3298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3199..3218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3452..3469
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3484..3624
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3550..3569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3710..3732
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3888..4766)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tacccatggg tcatcatcac
      361 catcaccaca agccagagac caagctcgga aaagatctgc aagtattcaa tcgaattgcg
      421 ggcgctaccc acttaattca ggggattgtg ctgctcgtga tcatgaatac cgagaccaca
      481 attccagtgg taacgcgctt ctttgatcag acatctgacg gtgaaattgc cccggtcagc
      541 aaaattctct ttgagttttc agttgcgaaa atcgcaccga tctttctcct gctttcagca
      601 tttgcccata ttttaatttc atcaccttct tatgttcgcc gttacgagca gaacatttcc
      661 aaggggatta acccagtgcg ctggtgggag tatgccttta gctcttcact gatgctcgta
      721 gttctgctga tgtttggtgg cttaatcgag ctttccacag ttgtctttgt cttcaccctt
      781 aacttcatca tgaacctcat ggggctagta atggagaaat acaaccaact aacccagacc
      841 acaaactggt tgccctttaa catcggcgtt ctcgccggaa ttgttccgtg gattatgggc
      901 ggactttact tctgggtttc cacaaacaat attgcagatg cgattccggt ctatgcccag
      961 ttcggatttc ttcttacctt cttattcttc aatagctttg cgataaatat gtggttgcaa
     1021 tataagaaag tcggaaagtg gagcaattac gcatacggcg agaatgccta catagttttg
     1081 agcctagttt cgaaatccgc cctcggctgg atcatcgtgc tcgggacaat ggggatttag
     1141 gtttaaacgg tctccagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     1201 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     1261 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     1321 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     1381 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     1441 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     1501 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     1561 ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta
     1621 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1681 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1741 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1801 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1861 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1921 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1981 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2041 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2101 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2161 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2221 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2281 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2341 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2401 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2461 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2521 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2581 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2641 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2701 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2761 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2821 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2881 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2941 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3001 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3061 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3121 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3181 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3241 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3301 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3361 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3421 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3481 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     3541 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3601 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3661 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3721 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     3781 gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa
     3841 accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta tgacaacttg
     3901 acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg
     3961 gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg
     4021 acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg
     4081 tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc
     4141 gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg
     4201 tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac
     4261 tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc
     4321 tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa
     4381 tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag
     4441 ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg
     4501 cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca
     4561 cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg
     4621 agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg
     4681 ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc
     4741 aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag ag
//
