LOCUS       sequence-365702-        5577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of small FRET-based near-infrared genetically-encoded 
            calcium indicator in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5577)
  AUTHORS   Matlashov ME, Vera J, Kasatkina LA, Khodakhah K, Verkhusha VV
  TITLE     Design and Initial Characterization of a Small Near-Infrared 
            Fluorescent Calcium Indicator.
  JOURNAL   Front Cell Dev Biol. 2022 Jun 29;10:880107. doi: 
            10.3389/fcell.2022.880107. eCollection 2022.
  PUBMED    35846350
REFERENCE   2  (bases 1 to 5577)
  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"; journalName: "Front Cell 
            Dev Biol."; date: "2022-06-29"; volume: "10:880107. doi"; pages: " 
            10.3389/fcell.2022.880107. eCollection 2022"
COMMENT     Sequence Label: pBAD-iGECInano
FEATURES             Location/Qualifiers
     source          1..5577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(155..1033)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1060..1344
                     /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     1267..1286
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1387..1404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3037..3054)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3207..3293
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3385..3412
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3431..3522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3523..3591,3592..4383)
                     /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(3741..3760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4554..5142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5043..5062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5296..5313
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5328..5468
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5394..5413)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5554..5576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc
       61 ggcatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
      121 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
      181 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
      241 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
      301 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
      361 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
      421 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
      481 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
      541 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
      601 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
      661 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
      721 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
      781 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
      841 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
      901 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
      961 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     1021 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     1081 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     1141 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     1201 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     1261 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     1321 cgcaactctc tactgtttct ccatacccgt tttttgggct aacaggagga attaaccatg
     1381 gggagccacc atcaccatca ccatggcaga tctgcagctg gtaccatggc caatctcgat
     1441 aaaatgctca atacaacagt gaccgaggtg cgcaagttcc tgcaagcgga cagagtgtgc
     1501 gtgttcaagt tcgaggaaga ttactccggc accgtctcgc acgaagccgt ggacgacaga
     1561 tggattagca tcctgaagac ccaggtgcag gacagatact tcatggaaac cagaggcgag
     1621 gaatacgtcc acggcagata ccaggccatc gccgacatct acacagccaa tctggtcgag
     1681 tgctacagag acctgctgat cgagtttcag gtgcgggcca ttctggctgt ccccatcctg
     1741 caaggcaaga agctgtgggg cctgctggtg gcccaccaac tggccggccc tcgggagtgg
     1801 cagacctggg aaatcgactt cctgaaacag caagctgtcg tcatgggaat tgctattcaa
     1861 cagccgccaa gcgaagagga actgagcgag tgcttcagaa tcttcgactt cgacggcaac
     1921 ggcttcatcg acagagagga gtttggcgac atcatcagac tgaccggcga gcagctgacc
     1981 gacgaggacg tggacgagat cttcggcgac tccgacaccg acaagaacgg cagaatcgat
     2041 ttcgacgagt tcctgaagat ggtggaaaac gtgcagccga cctgcgacga tgagccgatc
     2101 catatccccg gtgccatcca accgcatgga ctgctgctcg ccctcgccgc cgacatgacg
     2161 atcgttgccg gcagcgacaa ccttcccgaa ctcaccggac tggcgatcgg cgccctgatc
     2221 ggccgctctg cggccgatgt cttcgactcg gagacgcaca accgtctgac gatcgccttg
     2281 gccgagcccg gggcggccgt cggagcaccg atcactgtcg gcttcacgat gcgaaaggac
     2341 gcaggcttca tcggctcctg gcatcgccat gatcagctca tcttcctgga gctggagcct
     2401 ccccagcggg acgtcgccga gccgcaggcg ttcttccgcc gcaccaacag cgccatccgc
     2461 cgcctgcagg ccgccgaaac cttggaaagc gcctgcgccg ccgcggcgca agaggtgcgg
     2521 aagattaccg gcttcgatcg ggtgatgatc tatcgcttcg cctccgactt cagcgggtcc
     2581 gtgatcgcag aggatcggtg cgccgaggtc gagtcaaaac taggcctgca ctatcctgcc
     2641 tcattcatcc cggcgcaggc ccgtcggctc tataccatca acccggtacg gatcattccc
     2701 gacatcaatt atcggccggt gccggtcacc ccagacctca atccggtcac cgggcggccg
     2761 attgatctta gcttcgccat cctgcgcagc gtctcgccca accatctgga gttcatgcgc
     2821 aacataggca tgcacggcac gatgtcgatc tcgattttgc gcggcgagcg actgtgggga
     2881 ttgatcgttt gccatcaccg aacgccgtac tacgtcgatc tcgatggccg ccaagcctgc
     2941 aagagggtcg ccgagaggct ggccactcag atcggcgtga tggaagagtg agaattcgaa
     3001 gcttggctgt tttggtggat gagagaagat tttcagcctg atacagatta aatcagaacg
     3061 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     3121 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     3181 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3241 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     3301 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     3361 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     3421 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3541 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3601 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3661 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3721 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3781 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3841 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3901 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3961 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4021 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4081 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4141 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4201 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4261 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4321 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4381 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4441 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4501 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4561 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4621 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4681 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4741 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4801 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4861 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4921 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4981 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5041 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5101 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5161 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5221 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5281 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     5341 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     5401 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     5461 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5521 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggt
//
