LOCUS       sequence-352857-        4807 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 4807)
  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 4807)
  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: HULAa36F11
FEATURES             Location/Qualifiers
     source          1..4807
                     /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(1208..1225)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1378..1464
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1556..1583
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1602..1693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1694..1762,1763..2554)
                     /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(1912..1931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2725..3313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3214..3233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3467..3484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3499..3639
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3565..3584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3725..3747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3903..4781)
                     /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 ttgagacgga aactaagttc aagaagctga agaattttaa tctgactatg
      421 gggattctgc acctgttcca gggaattctg atggtggtgc tgagcaatga cttcgcactc
      481 cctgtgacga ggagctatct tgcggctgaa tacccgaccg gaactaccgg cggtatgcct
      541 gctctcataa ccgtgagcga gacgctgttc gaagtctgga tagggcctct cgtcgccctg
      601 ttcctcttca tctccgcagc ggcacatatc ctgatatcca ctgtccttta caagaaatat
      661 attgccggct taatgaatca ccagaacaga tacaggtggt acgagtacgc cataagctcc
      721 tccctgatga ttgtcgttat cagtatgctt gttggcattt cggacatcgg aactttgatg
      781 ctggccttct tcctcaacat gatgatgatt cttttcggat tgctgatgga gaccatgaac
      841 gaaggtcgca ggaagcttga ttggtcgcct ttctggttcg gatgcattgc cgggttcata
      901 ccctgggtgg tcatattcgt ttggctgttc ggagccggag gctcgggcgg aggtccgccg
      961 gatttcgttt actggatatt cctcagcatg gcgattttct tcaacagctt cgcggtaaac
     1021 atgtggctcc agtacaagaa gaaaggcaag tgggccgatt acctttacgg cgagcgtatg
     1081 tatgttatac tgagccttgt tgcaaaatca ctgctggcat ggcaggtttt tgcgggcacg
     1141 ctcaggccgg cataggttta aacggtctcc agcttggctg ttttggcgga tgagagaaga
     1201 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     1261 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1321 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     1381 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1441 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     1501 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     1561 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     1621 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1681 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1741 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1801 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1861 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1921 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1981 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2041 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2101 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2161 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2221 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3421 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3481 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3541 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3601 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3661 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3721 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3781 cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca aatggacgaa
     3841 gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc
     3901 aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg
     3961 ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa
     4021 ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc
     4081 tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa
     4141 agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa
     4201 ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc
     4261 ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga
     4321 tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca
     4381 aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca
     4441 aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac
     4501 tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg
     4561 aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc
     4621 tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa
     4681 aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac
     4741 gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca
     4801 ttcagag
//
