LOCUS       sequence-138902-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish transgenic construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  AUTHORS   He J, Wang Y, Missinato MA, Onuoha E, Perkins LA, Watkins SC, St 
            Croix CM, Tsang M, Bruchez MP
  TITLE     A genetically targetable near-infrared photosensitizer.
  JOURNAL   Nat Methods. 2016 Mar;13(3):263-268. doi: 10.1038/nmeth.3735. Epub 
            2016 Jan 25.
  PUBMED    26808669
REFERENCE   2  (bases 1 to 5612)
  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: 
            "10.1038/nmeth.3735"; journalName: "Nat Methods"; date: "2016-03"; 
            volume: "13"; issue: "3"; pages: "263-268"
COMMENT     Sequence Label: pISceI-myl7:MBIC5-mCer3
FEATURES             Location/Qualifiers
     source          1..5612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             127..846
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     CDS             join(904..906,907..909,910..1623)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(949..970)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1210..1229)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1557..1578
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1661..1795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1685..1704)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1739..1758
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1908..1928)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1908..1926)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1947..1963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1947..1963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1960..1982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1971..1987
                     /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)."
     promoter        complement(join(1995..2001,2002..2019,2020..2025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2040..2061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2178..2195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2349..2937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2429..2448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3108..3899,3900..3968))
                     /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     3731..3750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3969..4073)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4099..4554)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4236..4257)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4448..4467
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4628..5612
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccggcttg gggatatcca ccatggaatt cggcctttct
      121 ggcctgcagg ccgtcgttac ccaagaacct agtgttaccg ttagcccagg tggtactgtt
      181 atacttactt gtggaagtgg tacgggtgcc gtcacatctg gtcattatgc aaattggttt
      241 caacaaaaac caggacaagc tccaagagct ttgatttttg atactgataa gaagtattct
      301 tggaccccag gtagattttc tggatctttg ctgggagcaa aggcagcttt gacaatatca
      361 gatgctcagc ctgaggacga agccgagtat tactgttctc ttagcgacgt ggatggctac
      421 ttgtttggcg gtggaacaca actgacggtt ctgtccggtg gtggcggctc tggtggcggt
      481 ggcagcggcg gtggtggttc cggaggcggc ggttctcagg ctgtggtgac tcaggagccg
      541 tcagtgactg tgtccccagg agggacagtc attctcactt gtggctccgg cactggagct
      601 gtcaccagtg gtcattatgc caactggttc cagcagaagc ctggccaagc ccccagggca
      661 cttatatttg acaccgacaa gaagtattcc tggacccctg gccgattctc aggctccctc
      721 cttggggcca aggctgccct gaccatctcg gatgcgcagc ctgaagatga ggctgagtat
      781 tactgttcgc tctccgacgt tgacggttat ctgttcggag gaggcaccca gctgaccgtc
      841 ctctccgcgg ccggtctggc ccatatggga ggcggtgggt ctggtggcgg tggcagcacc
      901 ggtatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      961 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1021 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1081 accctcgtga ccaccctgag ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1141 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1201 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1261 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1321 cacaagctgg agtacaacgc catccacggc aacgtctata tcaccgccga caagcagaag
     1381 aacggcatca aggccaactt cggcctcaac tgcaacatcg aggacggcag cgtgcagctc
     1441 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1501 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1561 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1621 taactcgagc ctctagaact atagtgagtc gtattacgta gatccagaca tgataagata
     1681 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1741 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1801 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     1861 cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
     1921 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2761 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4081 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4141 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4201 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4261 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4321 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4381 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4441 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4501 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4561 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4621 gccagtcgac catagccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     4681 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     4741 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     4801 ggacttggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     4861 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     4921 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     4981 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     5041 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     5101 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     5161 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     5221 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     5281 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     5341 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     5401 ggcaagtaca ttactattgg aaggacgcca gggtacattg gcagtactcc cattgacgtc
     5461 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     5521 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
     5581 aggtctatat aagcaatgct cgtttaggga ac
//
