LOCUS       sequence-352862-        4822 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 4822)
  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 4822)
  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: HULAa45C8S
FEATURES             Location/Qualifiers
     source          1..4822
                     /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(1223..1240)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1393..1479
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1571..1598
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1617..1708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1709..1777,1778..2569)
                     /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(1927..1946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2740..3328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3229..3248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3482..3499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3514..3654
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3580..3599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3740..3762
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3918..4796)
                     /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 ccaagaaaat ccgcaccatc gacgaacaaa tcaatcgtct tcgcatctac
      421 aacgtcgttg ccggcgccat tcacctgggc caggcgctcg cattcggata cgcgctgaca
      481 ctcatcagca cgcccgtctt gttcccagtg accgttgact acatgactgg ccctcccgga
      541 gtcccactcc ccacggaacg cgtcacactc ttcgaaatcg atctcggcat cggcgttgtt
      601 gccttcttgt tgatgtcagc gtttttccac ttcctcatct cgcttccccg ggtgttcacg
      661 cgctacgcga atggccttcg ggcaaaccac aactatttcc gttggacgga atactcgctg
      721 agctcgtcgg tgatgatttg gctcatcgcc cagctaacgg gcagcactga cgtcggtgca
      781 ctcttcggca ttttcgccgt caacgcgtcg atgattatgt tcggcgcttt gcaggagaag
      841 tacgagaagc ccggcaacgg taacttcttg cccttcgttt ttggttccat gacgggaatc
      901 gtgccctgga tcatcatcct gatttacaca cttcagcccg gctcagagtc cgcagctgag
      961 gttcccgggt tcgtcatcgg catcatcgtg tcgctattcc tgttcttcaa caccttcgcc
     1021 gtcaaccagg tgcttcaata caagcaaatc ggcggttggc gtgactacgt tcggggtgag
     1081 cgcatgtaca tcacactgag ccttgtggcg aagagcgcgc tcgcatggca agtgttttcc
     1141 ggtgcactcg ttccgctgtt tgtcgactaa gtttaaacgg tctccagctt ggctgttttg
     1201 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     1261 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     1321 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     1381 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1441 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     1501 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     1561 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     1621 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1681 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1741 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1801 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1861 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1921 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     1981 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2041 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2101 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2161 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2221 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2281 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2341 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2401 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2461 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2521 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2581 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2641 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2701 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2761 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2821 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2881 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2941 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3001 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3061 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3121 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3181 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3241 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3301 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3361 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3421 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3481 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3541 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3601 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3661 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3721 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3781 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc
     3841 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt
     3901 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa
     3961 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag
     4021 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc
     4081 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct
     4141 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
     4201 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
     4261 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac
     4321 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
     4381 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
     4441 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
     4501 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
     4561 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
     4621 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
     4681 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
     4741 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
     4801 ttcatactcc cgccattcag ag
//
