LOCUS       sequence-352853-        4825 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 4825)
  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 4825)
  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: HULAa2F4
FEATURES             Location/Qualifiers
     source          1..4825
                     /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(1226..1243)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1396..1482
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1574..1601
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1620..1711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1712..1780,1781..2572)
                     /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(1930..1949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2743..3331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3232..3251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3485..3502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3517..3657
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3583..3602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3743..3765
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3921..4799)
                     /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 catcaccacg ctaaagttga aagaactcca gatcaataca tttcaagatt gcgctcctac
      421 aacgtcgtag ctgggtttct acacctagct caggctgcgg gacttaccta cgtcttgacg
      481 ttacttgaaa cacaaatttt gtttccggtc acaattgact atccgacagg ccctccggga
      541 gtgccggttc cgccagagcg agtcgaactt tttgacatca acatcggagc gggagtcatc
      601 ggcttcttac tactttccgc tttctttcat ttcctgattg catcgccgct gtttttccag
      661 cgctataaaa atggcctaaa acaaaatcac aattacttcc gctgggttga gtattcgctc
      721 tcatcctcag tgatgatctg gctgattgct cagctgacag gcgtatcaga ctttgcagcg
      781 ctggccggaa tctttgccgt caacgcttca atgattttct ttggtgcttt gcaggagaag
      841 tatgagactc caggtaacgg caagggactt ccattcatca tgggaagcat gaccggtatc
      901 gttccctggc tgatcatcat cgtctacaca attcagcctg gtagcgaatc agctgccgag
      961 attcctggct tcgtttacgg aattatcgtt gcgctattca tcttcttcaa cacctttgcc
     1021 ttcaaccagg cattccaata caaaaagatt ggaccttgga gaaactacct atacggcgaa
     1081 gcaacctaca tcacgctaag cctggtagca aagtcgatat tggcttggca ggtattctcg
     1141 ggagcaatca tcccggcgat tgccagcaca taagtttaaa cggtctccag cttggctgtt
     1201 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1261 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1321 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     1381 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1441 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     1501 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     1561 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     1621 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1681 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1741 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1801 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1861 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1921 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     1981 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2041 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2101 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2161 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2221 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2281 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2341 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2401 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2461 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2521 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2581 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2881 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3421 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3481 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3541 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     3601 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3661 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3721 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3781 ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt
     3841 gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca
     3901 attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca
     3961 caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa
     4021 tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg
     4081 ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc
     4141 cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg
     4201 acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg
     4261 cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt
     4321 aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg
     4381 gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga
     4441 aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc
     4501 ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga
     4561 tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc
     4621 tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc
     4681 agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc
     4741 accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata
     4801 cttttcatac tcccgccatt cagag
//
