LOCUS       sequence-352858-        4780 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 4780)
  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 4780)
  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: HULAb132A11
FEATURES             Location/Qualifiers
     source          1..4780
                     /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(1181..1198)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1351..1437
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1529..1556
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1575..1666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1667..1735,1736..2527)
                     /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(1885..1904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2698..3286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3187..3206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3440..3457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3472..3612
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3538..3557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3698..3720
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3876..4754)
                     /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 ctaaagagat caatgattca aagctgcgcc gatacaacat tattgccggc
      421 gtattccatc tagttcagat gattgcagtc ttggctcttg ccaatgactt cactcttcca
      481 attgttgctc gttatatgtc cggccctccg ggaagcacct tcgcagagcc aatcaccttg
      541 ctaaacactc cagtcggaat cggcgtcgct ctattccttg gtctttcagc tttcttccac
      601 ttcttagtgg cttcaccaca gttcttctca cgctacagcg caggccttgc acaaaaccgc
      661 aattacttcc gctgggttga gtactcaatc agctcatctg taatgatcgt gttgatcgct
      721 cagatctgcg gagtctcaga tgtcgtcgcg attatttcaa tctttggcgt gaacgcttca
      781 atgatcttgt ttggttggct acaagagaag tacgagacac caggaaatgg cggatggatt
      841 cctttcatct ttggttgcat cacaggaatc atcccttggc tagctttggt tttctatgtt
      901 ttctcaatcg gcggtccatc agaggcatca gctccggcat ttgtttacgg aatcgttttc
      961 tcactcttcg tacttttcaa tacctttgca gtcgttcaat atctgcagta caagaaggtt
     1021 ggaaagtggt ctgactacct aagtggagaa aagacataca taactctctc tttgatcgct
     1081 aaatcagcac ttgcttggca ggtttttgtc ggcactttga ttgagtaagt ttaaacggtc
     1141 tccagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     1201 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     1261 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     1321 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     1381 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     1441 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     1501 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     1561 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1621 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1681 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1741 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1801 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1861 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1921 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1981 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2041 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2101 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2161 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2221 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2281 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2341 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2401 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2461 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2521 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2581 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2641 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2821 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     3541 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     3601 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3661 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3721 ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag
     3781 cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
     3841 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
     3901 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
     3961 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
     4021 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
     4081 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
     4141 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
     4201 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
     4261 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
     4321 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
     4381 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
     4441 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
     4501 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
     4561 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
     4621 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
     4681 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
     4741 tgcgcttcag ccatactttt catactcccg ccattcagag
//
