LOCUS       sequence-311673-        8499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized near-infrared fluorescent calcium indicator 
            NIR-GECO2 with codon-optimized heme-oxygenase in C.elegans 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8499)
  AUTHORS   Qian Y, Cosio DMO, Piatkevich KD, Aufmkolk S, Su WC, Celiker OT, 
            Schohl A, Murdock MH, Aggarwal A, Chang YF, Wiseman PW, Ruthazer ES,
            Boyden ES, Campbell RE
  TITLE     Improved genetically encoded near-infrared fluorescent calcium ion 
            indicators for in vivo imaging.
  JOURNAL   PLoS Biol. 2020 Nov 24;18(11):e3000965. doi: 
            10.1371/journal.pbio.3000965. eCollection 2020 Nov.
  PUBMED    33232322
REFERENCE   2  (bases 1 to 8499)
  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: "PLoS 
            Biol."; date: "2020-11-24"; volume: "18(11):e3000965. doi"; pages: "
            10.1371/journal.pbio.3000965. eCollection 2020 Nov"
COMMENT     Sequence Label: pSF11-wNIR-GECO2-T2A-HO1
FEATURES             Location/Qualifiers
     source          1..8499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4274..4285
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4277..4330
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(6016..6038)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6138..6157
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6239..6257)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6325..6429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6430..6498,6499..7290)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6648..6667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7461..8049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7950..7969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8203..8220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8337..8358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8373..8378,8379..8396,8397..8403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8411..8427
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8416..8438
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8435..8451
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8435..8451
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggatccatag gtatgcctgt cattctctat ttttaaacgt tcctgcagca gtagcacaac
       61 agagccagca gttttcctct tcttctccat cttcttctgc ttcttcctct cttcccaaaa
      121 aaaaaaaaaa aaaaaaanrw gagggggctc mctgcccccc cccccctccc ccccccctct
      181 tctccttgaa gctcatccag acgtcgctgt cctcctcctc ctcgtccccg gggcgctgtg
      241 tgtgtgtgtg tgtgctccgt gagcatgtag cagcttcagc ccatttaaag gcgcacactc
      301 tctgcatctc tgcgtgtctc gacactcgtt ttttctcttc ttttctgctg cacacacaca
      361 cacacacaca cacacgctca tgccagagca cttcttgctc cggggggctc cgcgcggagc
      421 ccgagaggac ctcctcctcc tttttcttct tcgcttttta gttcttcttg cctcggtctc
      481 gccccttgtt ttgctctcca tgtagtatat acatctgggt gtgtgtgtgt gtgtgcacct
      541 ttggagcatt ttttgaacct ttccagaact ttggagcaac attttcagct tgttcagtgc
      601 actttttagg ctgaaaatcc tatttttagg ttttcgaaag tttttttgga gcatttttcg
      661 tgcaatttag gtgcaaaatt aaaaaaaagg gttttgtagt gaattaaaat ttttttgaaa
      721 ctttcgcctt tttaaaaaaa tgtttctaat atctttaaaa attggaatat tttggaaaac
      781 aatttgtttt caattggcaa tttattaatt ctgagcttct gagcgtaaat ttttttttat
      841 tttggagcaa ttctgagatg tcttagtgca tttttcgggc atacataaaa aaaaatttat
      901 cttaataata aaaaccttaa atttaggctt caccgtctgg aaattcaatt ttttcagctg
      961 aaaaactttc aaaaaaaaaa aaattaaatt ttcacattta ttttctctgg aattttggag
     1021 caatttcaaa gctgtcttag tgcattttta gtacacaaat attttttcat ttttcttaaa
     1081 ttttaataat ctaagcttaa ataaaggctt caccgtctga aaatgcaact tttttcagat
     1141 gaaatattaa aaaaaaaacc tcttttatta attttgtgct ccaaactttc ccacggtacg
     1201 caaatcatgt aatatgtact tttaaacatt atttttccaa tcaaaaatat ccaacagctt
     1261 ttccttacaa tttgaaaatt tttattaaaa tttcaaaaaa attttttttt tttttaattt
     1321 cgaatttttt taaacttgtt gtaacttttt tccctgtcgg cgccaattaa caaaatacgt
     1381 agaattttgc gcttattgtt ttgttagtta acaaattttt gacaactttc ttatcagctg
     1441 agatttaggt gtttcagttc gcaatctgcg tctctccttg ctctcctcta ttttaaaacc
     1501 cccataactt tgttcttgtg agagttatca aaaaattgtc aactaacaaa atatagagca
     1561 caaaaatatg cacattttgt atgttgacaa ttttttgata ccttttttag ttttcgagat
     1621 atcttagttt tagtttagct gtaccttttt ctggcaaaaa acttccaaat ttcggcaatt
     1681 tttcggcaaa tttcccggca aatttcgcca agtacatcgg gctctcagac aagtggtctg
     1741 tgagtggaag tcgataaaag tagtttacgt gctctccgtc cccccgcttc ttctccgtct
     1801 ctgcttctat ttctccattg actagtccca ctacttttta cggggcggag ggctctcttg
     1861 cactccagct tcttctcttt ttgtgctctt ttttgcccgt atgtgggtgc tcatttattt
     1921 ttctttggtt ttaaaataaa ttcggtttat taggctactc gtaaaaatga agaaaaaact
     1981 tcaaattcaa acgacattaa aaaaaaattt aataaaaatt ttttgaattg ctcaaaagcc
     2041 ccacctatat atatatatag ctccgcctac ttctcgcaca gaccccgccc actttttgac
     2101 tccgccccct tattggcaaa agttcttgaa aaaaataaaa ataaaaattc aaattgaaac
     2161 aatttttttt taatgaagta gtttttttct aaattgatga aaatttctaa acagtttaca
     2221 agcaccgccc actttttgac tccgcccctt tgttggaaaa attcttgtaa aaaaaaataa
     2281 aaaataaaag cttctctttc aaacgatttt tagttggtct ttgtatgaaa agtttttttc
     2341 ctaaaataga aatgtctgaa taatttgcaa gctccgccct tttcttaccc agactccgcc
     2401 cctttattga ctaatttttt tttgtccatt tacctgaagt tcaaaaaaaa aatttacagt
     2461 ttcatagagt aataattttt caaaaatctc tttttccaat gtttaatgtg tcattataaa
     2521 cccgcccagt tttaagctcc gcccatttag gccccgccca tttatgaccc aaatattttt
     2581 ttctctattc acccgtggat ttccctccaa caagttaccc caaaaataaa aataaaaatt
     2641 cctgcaggcg cccaccggga tccggcgcgc ctctagagga tccccgggat tggccaaagg
     2701 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
     2761 ggctagcgtc gacggtacca tgtcggtgcc attaaccacc tctgccttcg gacatgcttt
     2821 cctcgctaac tgcgagcgtg agcaaatcca cttagcaggc tcaattcaac cgcatggcat
     2881 acttttggcg gttaaagaac ctgacaatgt tgtgatccaa gctagtataa atgcagccga
     2941 gtttttaaac accaatttcg tagtgggccg acctcttaga gatttgggtg gagacctccc
     3001 gttacaaata ctcccgcatt tgaacggtcc gcttcatctc gcgccaatga ctttgcgatg
     3061 cactgtgggt tccccaccaa gacgtgtaga ttgcacaata catcgtccgt caaatggagg
     3121 acttattgta gaattggagc cggcaacgaa agcgacgaat atcgctccgg ccttagttgg
     3181 agcgctccac agaataacgt catcgtcctc cttaatgggt ttgtgcgacg aaacagccac
     3241 catatttcgt gagattacgg gcttcgaccg agtaatggtg atgcgactcg gcgccctcga
     3301 tgaccttacc gaagaacaaa tcgcggagat taaagaggct ttttcgcttt ttgacaagga
     3361 tggagacggt accattacca ccaaggagct tggaaccgtc ttccgtagtt tgggccagaa
     3421 tccgacggag gccgaattgc aagacatgat caacgaggta gacgcagacg gtgatggcat
     3481 ctttgacttc ccggagtttc ttacaatgat ggcgcgtaaa atgaacgata cagactcgga
     3541 ggaagagatc cgaggtgcat ttcgtgtatt cgataaagac ggaaacggct acataggcgc
     3601 cgcagaatta cgacacgtaa tgacagacct tggtgagaag cttacagacg aagaggtgga
     3661 cgagatgatt cgagtcgccg ataatgacgg cgatggtcag gtaaactatg aggagtttgt
     3721 gcaaatgatg accgcaaaag gtggcggcgg atctgttgac tcaagtagac gtaaatggaa
     3781 caaggcggga cacgccgttc gtgccatcgg ccgtttgagt tccagaagac atgacctttt
     3841 gtcagaatgt cgtcgagcag acctggaagc atttctcgga aaccgatacc cggcttccac
     3901 aatccctcaa attgcaagaa gactgtacga gcttaacaga gtgcgtttgc tggtggatgt
     3961 gaattatacc ccagtaccac ttgagccacg tatatcccct ttaaacggtc gagacctcga
     4021 catgtcatta tcgtgtctgc gaagtatgtc tcctattcac cagaagtata tgcaggatat
     4081 gggcgtgggt gctacgcttg tttgttcgtt gatggtaagt ggcagattgt ggggacttat
     4141 tgtatgtcac cactacgagc caagatatgt cccatcgcac atccgtgccg ccggcgaggc
     4201 gctcgctgaa gcgtgtgcta atcgtatcgc cacacttgag tcattcgcac agtcgcaatc
     4261 aaagggttac gatattgaag gaagaggaag tttattgacc tgtggagacg tggaagaaaa
     4321 tccaggaccg atggaacgtc ctcaacctga ttccatgcct caggatttat ccgaggcctt
     4381 gaaagaagcc acaaaagaag tgcataccca agcagaaaac gcagaattca tgcgaaactt
     4441 ccaaaagggt caggtaaccc gagacggatt caagctcgtc atggcttccc tgtaccatat
     4501 atacgtggcc ctcgaggaag aaattgagag aaacaaagag tcgccagttt ttgcgccagt
     4561 gtacttcccg gaagagctgc acagaaaagc tgcattggaa caggacctgg cattttggta
     4621 tggaccaaga tggcaggaag tcattccgta cacacctgca atgcaaagat atgtgaagag
     4681 attgcatgag gtcggccgaa cagagcctga gctgttagta gcgcatgcct atacccgtta
     4741 tcttggcgac ttgtcaggtg gacaagtatt gaagaaaatt gcccaaaaag ctctggattt
     4801 accaagttct ggagaaggat tagcattttt tacatttccg aatatagcaa gtgccactaa
     4861 attcaagcaa ctctatcgta gtagaatgaa ctctctcgag atgacaccgg ccgtcagaca
     4921 gcgagtcata gaagaagcga agacggcctt tttgctgaat atacaattgt ttgaggagct
     4981 ccaggagctg ctgactcacg ataccaaaga ccagtcaccg agtcgagcgc caggcttgag
     5041 acaacgtgca tctaacaagg tgcaggattc cgcgcctgtc gagacacctc gtggcaagcc
     5101 gcctttaaac acacgatcac aagcaccact gttgcgttgg gtattgacgt tgtcgttcct
     5161 cgtggcaact gtggccgttg gtctgtacgc catgtaagag ctccgcatcg gccgctgtca
     5221 tcagatcgcc atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc
     5281 ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca aaaaacttct
     5341 cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat
     5401 tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt
     5461 aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta tgttttttac
     5521 ttctgataaa tttttttgaa acatcataga aaaaaccgca cacaaaatac cttatcatat
     5581 gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc ctctcatgac
     5641 gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt caatcaagtg
     5701 aaagtttatg aaattaattt tcctgctttt gctttttggg gtttccccta ttgtttgtca
     5761 agatttcgag gacggcgttt ttcttgctaa aatcacaagt attgatgagc acgatgcaag
     5821 aaagatcgga agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa gttgataatt
     5881 tgaaagtgga gtagtgtcta tggggttttt gccttaaatg acagaataca ttcccaatat
     5941 accaaacata actgtttcta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg
     6001 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6061 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6121 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6181 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt aattaagggc
     6241 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6421 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6481 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6541 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6601 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6661 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6721 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6781 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6841 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6901 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6961 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7021 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7081 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7141 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7201 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7261 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7321 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7381 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7441 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7501 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7561 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7621 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7681 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7741 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7801 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7861 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7921 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7981 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8041 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8101 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8161 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8221 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8281 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8341 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8401 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8461 gccaagcttg catgcgcggc cgcactgact gggccggcc
//
