LOCUS       sequence-352860-        4783 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 4783)
  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 4783)
  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: HULAa30F3
FEATURES             Location/Qualifiers
     source          1..4783
                     /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(1184..1201)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1354..1440
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1532..1559
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1578..1669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1670..1738,1739..2530)
                     /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(1888..1907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2701..3289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3190..3209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3443..3460
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3475..3615
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3541..3560)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3701..3723
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3879..4757)
                     /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 ctactgattc cgcgacggaa aagaaactca aggggctccg gacctggaac
      421 attgtggtgg gcctgatact ggcggtgcag gcggtgctga ttgccgtgct gaccaacagc
      481 ttcgcgctgc cggtgaccgc gacgtttatg gaggggccgc cggggaccgc gccggcgctg
      541 cagcatctgt ttaatatcca gacggggtgg ggcgtgttca tcttcatggc gatatccgcc
      601 ggcgcgctgc tgctgattgc ctccccgatg gttttctcct ggtacaagcg caatctgctg
      661 gaaagccgga actacgggcg ctggattgag tatttcttca gttcctccgt catgatcgtt
      721 ctgatatccc aaatctgcgg catcagcgat attgccgcgc tactggcgat attcggcatc
      781 aacgcgtcca tgatactctt cggcgcattg caggagaagt atgagaagcc gggcaagccg
      841 aacctgcttt cgttctggtt tggcagcttc gcggggataa ttccctggat tgccgtttta
      901 atctacgtgg tatcccccgg cgtggcggcg gcgccgcccg cgtttgtgta cggaattgta
      961 gtggcgctgt tcgtgttctt caactgcttt gcggtgaaca tgattctcca gtacaagcag
     1021 atagggccgt ggaaagacta cctgttcggc gagaaggtct atataatact gagcctgacc
     1081 gcgaaatccc tgctggcatg gatggttttc gcgaacgtac tggtcctgta agtttaaacg
     1141 gtctccagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     1201 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     1261 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     1321 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     1381 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1441 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     1501 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     1561 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     1921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2221 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2281 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2341 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2401 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2521 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2581 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2821 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3361 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3421 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3481 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     3541 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3601 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3661 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3721 aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg
     3781 aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
     3841 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
     3901 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     3961 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     4021 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     4081 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     4141 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     4201 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     4261 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     4321 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     4381 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     4441 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     4501 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     4561 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     4621 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     4681 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     4741 ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
