LOCUS       sequence-349041-        4793 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 4793)
  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 4793)
  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: EINA62G7
FEATURES             Location/Qualifiers
     source          1..4793
                     /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        1588..1679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1680..1748,1749..2540)
                     /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(1898..1917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2711..3299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3200..3219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3453..3470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3485..3625
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3551..3570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3711..3733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3889..4767)
                     /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 agttgcgata atcgcaccga tctttctcct gctttcagca
      601 tttgcccaca 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 catcggtgtt ctcgctggaa ttgttccgtg gattatggga
      901 ggactttact tctgggtttc cacaaacaat attgcagatg cgattccggt ctatgcccag
      961 ttcggttttc ttctaacctt cttattcttc aatagctttg cgataaatat gtggttgcaa
     1021 tataagaaag tcggaaagtg gagcaattac gcatacggcg agaaggccta catagttttg
     1081 agcctagttt cgaaatccgc cctcggctgg atcatcgtgc tcggaacaat ggggatttaa
     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 aactcttttt gtttattttt ctaaatacat tcaaatatgt
     1621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1921 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2221 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2821 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3361 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3421 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3481 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     3541 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     3601 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     3661 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3721 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     3781 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     3841 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     3901 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     3961 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     4021 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     4081 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     4141 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     4201 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     4261 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     4321 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     4381 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     4441 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     4501 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     4561 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     4621 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     4681 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     4741 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
