LOCUS       sequence-460022-        6574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear green fluorescent protein in zebrafish 
            hepatocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6574)
  AUTHORS   Eski SE, Mi J, Pozo-Morales M, Hovhannisyan GG, Perazzolo C, Manco 
            R, Ez-Zammoury I, Barbhaya D, Lefort A, Libert F, Marini F, Gurzov 
            EN, Andersson O, Singh SP
  TITLE     Cholangiocytes contribute to hepatocyte regeneration after partial 
            liver injury during growth spurt in zebrafish.
  JOURNAL   Nat Commun. 2025 Jun 6;16(1):5260. doi: 10.1038/s41467-025-60334-y.
  PUBMED    40480975
REFERENCE   2  (bases 1 to 6574)
  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: "Nat 
            Commun."; date: "2025-06-6"; volume: "16(1):5260. doi"; pages: " 
            10.1038/s41467-025-60334-y"
COMMENT     Sequence Label: fabp10a:H2B-mGreenLantern
FEATURES             Location/Qualifiers
     source          1..6574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..34)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(14..32)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(65..82)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(65..81)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(74..96)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(222..677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(359..380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     571..590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        704..808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(809..877,878..1669)
                     /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(1027..1046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1840..2428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2329..2348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2582..2599
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2716..2737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2752..2757,2758..2775,2776..2782)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2790..2806
                     /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     2795..2817
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2814..2830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2814..2830
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2876..2895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2876..2894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5122..5499
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     regulatory      5578..5587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5584..5586,5587..5589,5590..6303)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(5629..5650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    6433..6554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6470..6489)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6524..6543
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ccagcttttg ttccctttag tgagggttaa ttgcgcgcta gggataacag ggtaatacgc
       61 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      121 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      181 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat
      241 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      301 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
      361 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
      421 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
      481 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
      541 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
      601 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      661 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      721 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      781 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      841 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      901 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      961 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1021 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1081 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1141 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1201 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1261 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1321 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1381 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1441 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1501 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1561 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1621 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1681 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1741 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1801 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1861 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1921 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1981 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2041 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2101 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2161 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2221 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2281 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2341 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2401 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2461 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2521 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2581 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2641 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     2701 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     2761 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     2821 cagctatgac catgattacg ccaagcgcgt tagggataac agggtaatgc gcgcgtaata
     2881 cgactcacta tagggcgaat tggagctctg cagaagagca tctctgaaca cacaacacgt
     2941 ccaaccttga ggcagatggg ctacagcagc agaagaccac accgggtgcc gctcctgtca
     3001 gctaagaaca ggaaactgag gctacaattc acacagactc accaaaactg gacgatatat
     3061 tattagcccc ccttagaatt tttcatttga taatattttt cttctggcga aagcctcatt
     3121 tgttttatat attatagaat aaaattagtt tttaatagtt tttatgccat tttaaggtca
     3181 atattattag cccctttaag ctattttttt tcgatagtct acagaacaaa ccatcggtat
     3241 acaatgactt gcctaattac cctaacctgc ctagttaccc taattaacct agttaagcct
     3301 ttaaatgtca ctttaagctg tatagaagtg tcttgaagaa tatctagtct aatattattg
     3361 actgtcatca tggcaaagat aaaataaatc agttattaaa actattatga ttagaaatgt
     3421 gctgaaacaa tctgctctcc gataaacaga aattgaacaa aataaacagg ggggctaata
     3481 aatttaaggg gttaaataat tctgattgca aaaaaaatga tgtctgcaaa actgttgcaa
     3541 ataatttatt tgtgttgatt ttaagcaaac aaattaaatt taataaaata caacttaatc
     3601 tgtttgttta aattcagccc taataaattg tttacagcca cttaacgtaa aaaaattgag
     3661 taaatccaag gaatcatctc tgaataattt ttttcagtgt atatatatat atatatattc
     3721 ttacaaaaca actcatttac tttagtttaa ttttcagggg caaaaaacta aagtaatcga
     3781 cgttgcttga ataaaaagtg taattaaggg aatgaggtaa catttaacca tgtgtcaatg
     3841 cagtttaaat atgccagtta gtggtatatg tttaaatggt aagctattca aaactttaaa
     3901 ctaacttaac cagccttttg ttgtcagact gaacagactt tccatctgca ttattagaga
     3961 ctaatctttg gctggatgaa tgattcatct gctgatattt cagaatagac agattgaggc
     4021 tgtttctaat atgattatgc aacctgaggg tgattatttg aagcaaactc cacagaccag
     4081 caggtcattg accgtcgtgt gttcaaacag agcagaaaca tttgcaaaac tggtctgaca
     4141 ggagaatcca gtccagcaca acacatatgc tgagcaaact gaatcaatcc tgcaggtcaa
     4201 ctctcgtgct ttaagtttat taagattatt ttatttattt attattttat ttatctattt
     4261 atttatttag ttgtttattt agcaattatt tgtttattta tttgtttatt tattcctgca
     4321 gatcatgcct tgtgcctttt tacatttaat ttaattttta atttaatttc cttttacttt
     4381 tttttatttt tttattttat tttattttac agtctgacaa atactgaact aaaaaccctc
     4441 agatcatgtc ttatgcattt matttnantt tatttatttt attatttatt ttatttyatt
     4501 rtttatttta ttttatttta ttttatttta cagtctgaca tactggacca ataaatctct
     4561 cagatcatgt ctcatgcatt ttacttttat tttattagaa ttagaaagat caaaggaaca
     4621 acttttaaaa tattaattct gtatcaaaat ctcttttgat acatttaatt gatttaaaaa
     4681 agcagttcac ccaagaaaca tttcctcaca gtcgaatggt tgtaaacttt tatgaattac
     4741 tttcacagaa aaagattttt ggaagaatat tggaaaaaaa gcagccattg acttccatag
     4801 taacaacaaa aaatactatg gaagtcaatg gctgtttttt caccattcgg tatcttcatt
     4861 ctggagcaga attttttggg tgatctgtcc ctttaagtcg tcaaatcctg gtgcaatatt
     4921 ccacatgcac tgacgagtct ttatttttgg tctgctactg ctgtgtgtgt gagggcattt
     4981 acctcatctt ctgctggagt tgatgaacgg tgggttgttc aaacagcagc aggtcattga
     5041 ctgaactcct ctcgatataa aagctgcaga tctgaagctg accttcactt tgtgttgatc
     5101 gctagcgaat tcgccaccat gatgccagag ccagcgaagt ctgctcccgc cccgaaaaag
     5161 ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc
     5221 cgcaaggaga gctattccat ctatgtgtac aaggttctga agcaggtcca ccctgacacc
     5281 ggcatttcgt ccaaggccat gggcatcatg aactcgtttg tgaacgacat tttcgagcgc
     5341 atcgcaggtg aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctccagg
     5401 gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc
     5461 gagggtacta aggccgtcac caagtacacc agcgctaagg gatctggggg tggaggctcc
     5521 ggcgggggtg gatctggagg tgggggcagc ggcggaggtg ggctagcgct accgcgggcc
     5581 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     5641 gacggcgacg taaacggcca caagttcagc gtccgcggcg agggcgaggg cgatgccacc
     5701 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     5761 accctcgtga ccaccttagg ctacggcgtg gcctgcttcg cccgctaccc cgaccacatg
     5821 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5881 tctttcaagg acgacggaac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5941 ctggtgaacc gcatcgtgct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     6001 cacaagctgg agtacaactt caacagccac aaggtctata tcacggccga caagcagaag
     6061 aacggcatca aggctaactt caagacccgc cacaacgttg aggacggcgg cgtgcagctc
     6121 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     6181 cactacctga gccatcagtc caaactgagc aaagacccca acgagaagcg cgatcacatg
     6241 gtcctgaagg agagggtgac cgccgccggg attacacatg acatggacga gctgtacaag
     6301 taattaatta agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     6361 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     6421 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6481 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6541 atcaatgtat cttaatcgag ggggggcccg gtac
//
