LOCUS       sequence-474490-        9502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mNeongreen tagged form of zebrafish nhsl1b. For in vitro 
            transcription (SP6) or expression through CMV..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9502)
  AUTHORS   Escot S, Hassanein Y, Elouin A, Torres-Paz J, Mellottee L, Ignace A,
            David NB
  TITLE     Nance-Horan-syndrome-like 1b controls mesodermal cell migration by 
            regulating protrusion and actin dynamics during zebrafish 
            gastrulation.
  JOURNAL   Commun Biol. 2025 Feb 28;8(1):328. doi: 10.1038/s42003-025-07689-6.
  PUBMED    40021913
REFERENCE   2  (bases 1 to 9502)
  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: "Commun 
            Biol."; date: "2025-02-28"; volume: "8(1):328. doi"; pages: " 
            10.1038/s42003-025-07689-6"
COMMENT     Sequence Label: pCS2+ Zf nhsl1b-mNeongreen
FEATURES             Location/Qualifiers
     source          1..9502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      5022..5031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5520..5654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5544..5563)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5598..5617
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5767..5787)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5767..5785)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5806..5822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5806..5822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5819..5841)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5830..5846
                     /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(5854..5860,5861..5878,5879..5884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5899..5920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6037..6054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6208..6796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6288..6307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6967..7758,7759..7827))
                     /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     7590..7609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7828..7932)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7958..8413)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8095..8116)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8307..8326
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8484..9468
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 atttaggtga cactatagaa tacaagctac ttgttctttt tgcaggatcc gccaccatgc
       61 cgtttcccga gagagccgtt gagcctcagc tgctgtgccg gttgaggggg agcgatggac
      121 ccgagaagag ctttataaca ccggacggtc gcaaagttcg caaaccggtt ttgttttcct
      181 ctctggagga agtttgttgt catacgttca ccagcatcct acatcagctg tccgatctat
      241 cccgacacgc cagtgacata ttcctgggaa ttgagactca agctgggcta atctcgcaca
      301 ggacatcgag gattcaagcg cgtttggaga ggatacagca caccgttcaa acgctcgatc
      361 caaaatcggt taaaatacct gtatctaatc tggatgagga gaaaaagtgg accgtacact
      421 acactgcccc atggcaccag caggagaacg tctttctgcc aggctccaga ccggcctgtg
      481 tggaagatct tcaccgtcag gccaaagtga acctgaaaac agctctcaga gaatgtgata
      541 aactgaggaa ggatggcttc cgcagctcgc agtactactc acaggggccc actttctctg
      601 gctcctcaag ctcccaacat gaggacgagg accttgaagg cgatgacgct gacaaaaagt
      661 ccacagcatc ttcaggagag gaagagaaag actcctgcca aatgagagcg caaagtccgg
      721 cacaggtgga aggagtcgag gttgatggac aaatgtcgag ctccaaagcc tcagtcctgc
      781 ccactccaga ggagaagatg aggcaacagg cacaggccgt tctcacagat attgtcccta
      841 tcaatgtcac ggcggctaaa gggcgtggag gagagcttcg accacagtcc atgtttatcc
      901 ctggacagta ttcaacactt gggcgagtca tgaatgggaa ttcaactctg cgtcaatctg
      961 taacaaaaga ttctggctgc cagactgagg cagtgaaaat cgtcccaccc tcagtaagga
     1021 gaatacgggc tcagaagggt caaggaattg ctgctcagat ggctggcatc tccacctcag
     1081 ccacaaacat ctcttctgtt tctgatggga gctccactgg aggttcgatt gttgcaatgg
     1141 caccacagtt cggcaatgat atccaacggt ttcacagctt gccaaggggt gcacgggtct
     1201 ctctaaatgc agagcccatt tacagtagca cccctttcag aaaggaagac tcctgtgcaa
     1261 aagcacctaa gcagattgga aaattacaag tagacaatac tgcagtgcac atgaggaatg
     1321 cccccagggt atgtactcaa gcccgcccaa aatctcagga ggtgcgagga tctcaaaggg
     1381 agtggggttc tgtctcgggt ccagcctgtg tagtttctcc acatgcagcc tactcaacct
     1441 cccttatacc taatgctact ctgtcatgtt ctgctgaggt ttttgctctt cacagcaaat
     1501 caagtcctgg ccagagccca ctttctgggt caccatacac taaagctaga cctcttagca
     1561 tggtttcagc tgtcaacagt gggagtacaa gtagtgtggg cacaggatct cacaccccag
     1621 atgcaggcat gaaagatacc tgcagtgaga ctggtcaatc tgatagcagt ttgcacagcc
     1681 acagtaccat tgccgcagga acattgtctt cagatgagca gtggatttac gatactcctg
     1741 ataatgcctt gcccagaagg acaatgactt ccagctgctc aacacccata aatcatctat
     1801 atagtagcct tgaacgctcc tcaaaaggca cagactctag ttctctgtac tctatggaca
     1861 atgatggcta ctacacttcc atgcatctgg attcaggcct taggtcaaga agccatggta
     1921 gcggccatgg tcatggcata ggaggaagag cagtaagaca cagtatgtac gagtgcttgg
     1981 gtgagcaaga ggatcgaaat agcttgtgca gcgaccactc gctatctcgt tccatttctc
     2041 tacgcaagcc taagaaacca ccccttccac ctgcacgcac agactctcta cggagaaagc
     2101 ctgggaaaaa cagctctccc tctgccaata ttgggcggtc aaatattagg aatggttctc
     2161 ttctcaatga atcattgatt gccactctcc aacagtcact tcagaatggg cttaaaggca
     2221 aaggatcctc cacgtcacca tctcatagcc cttgtagtga ctatgaggac ccctggatgc
     2281 ttcgtccaag gagccagagc agcatcagtg caagtagtag tggtgtatca gctacaggaa
     2341 tggctcatgt atactccatc tgtcccgtca caccatctca tagtgacaca agtagcctcc
     2401 gttctgatta cgccgactcc tggggctact acatggaata ccctcgtcca tctggagatc
     2461 agacacagtc accatgtacc aattcacact ctgctggaca agtgtctgag atggcaaatg
     2521 gaagaggccc tctcaatggt aaccaggcta atcttcctgc tgttcaggaa ggaagtgaca
     2581 tgtctgctaa gcccaggaca ggcacctctt caccggacag ggtacatcgg ttgacttctc
     2641 catcaagtgg atactcaagt caatcaaaca ctccaactgc tggtactccc gttccctcct
     2701 tcatgagatg catgtcccca tcaggcaagc ccaaaccaag agtgcctgaa agaaagtcat
     2761 ccttgctttc atctgtatct atatcctcct cttccacctc tctttcatcc aacacctctg
     2821 attctaatag gagcaatatt ccacctcctc ctcctcttcc agctgcccct gtggtacttg
     2881 tacctcttaa tcctccatcc tttcctcctc cccttccacc cccaagccct gtgtgcactg
     2941 aagaccagaa atttccacct ccgccacctc ctttacccac aactccccat cagcaagcat
     3001 ccatgaaccc tcttcaaaac aattcctccc ctgaatttcc accccctcca cctccagaag
     3061 tgttgaccga ccctgcctgt tcaggcttta atagttcctt cagccctcca ccacctcccc
     3121 ctccaccacc actgcctact tattctcctt cccaaaatca acatttaccc cacatgacgc
     3181 cacaaacacc ttcttctgcc tccttgaaag aaattaaggg ctgtcttaaa ccagtgaact
     3241 tagaaggaag accagtgttg cctcaatctg aggaacctaa taaggttagc atgcctttga
     3301 ttaccccatt tgcactgcag agtgtgcagc ttcggccagc aaaacgtcca gaaaacaatg
     3361 aagttagcag gacagacaga ccccaaacac cagaaaaaac tcaccccaca gttcctcaga
     3421 tttcttgtat atctccaaat gtaagccttt catctacaga gcaaaaaccc tcaatccagc
     3481 atagccagga tggccttgtt gaagcacaaa ctgactttag tgtaaaatcc aattctgatc
     3541 caacatcctt ccttacaaat gggcccatag atcttggtga ggccacagat gaagtagatg
     3601 gtttaaagac tttcctctca acacaagtgg aaaagtcact aaactctaca cccaagaaaa
     3661 agcctccaat gatttccaaa aagcccaagt tttctcttac tttctcctct gtggatcatg
     3721 taagggattt tgtgagtgct gaaggtctgg accccatacc tgtgctggaa aaagaggaag
     3781 agacaactga gaaaagtagc tttgacattt ccgtacctcc tgtagagata agagacattt
     3841 ctcccacaaa tggtgaatct caggagttct cacaatcctc tggtaccact attcttgatg
     3901 tagacaagag cacctctgaa gcagaggata aagaagaaag agatgacgaa gatataacca
     3961 gcagcacagg atcgttcagt tctaaagatg atgaaaatgg tgatgtgttt gaatccagca
     4021 cgttggacgt ggctcaatca cccagcatca acggtgacag ctgtggggac atggtgaccc
     4081 ccacacggcc ccgcaccaca gaggacctgt ttgcagccat tcacaggtcg aagagaaaag
     4141 tcctggggcg caaggaatca gaagaggatc gctcccgggg tcctctatct ccaccggtca
     4201 ccccgacagc accgagcccc agcttgacct cctctctgcc ccggcaatca ggctccatcc
     4261 agcgcagcct ccgcaaatct gcaaccagca gtgacacctt caaggccctt ctgctaaaga
     4321 aaggcagtcg ctctgaaagc agtttccgca tgtctgcaac cgagatgctg cgcagcaccg
     4381 acccgcgctt tcaacgcacc cgctccctcg actcctcctt tgatccggcg tctccaacat
     4441 ccgagagccc ctgctcttct ccaggccgaa gcaagagggt ggcagatgac tgggcgcgca
     4501 atgagggaat gttttcaact tcgccatctt tgataggccc aaaatatgga cgatcccgaa
     4561 cgccaccatc tgctgctagc agcaaataca acgcccgctg ccgcatcctc agcagcccca
     4621 tgactgtcat ctgcgagagg gatggagaac taaccgactg tgaagacccc gtttctccgt
     4681 caaacacacc ccttcccatc tctcaggact caaacagcac tttatgtgag cagagcagca
     4741 gtaccctcct gtggccaccg ggatccatgg tgtctaaggg cgaagaggac aacatggcca
     4801 gcctgcctgc tacacacgag ctgcacatct tcggatctat caacggcgtg gacttcgaca
     4861 tggtcggaca aggcacaggc aaccctaacg acggatacga ggaactgaac ctgaaaagca
     4921 ccaagggcga cctgcagttc agcccatgga ttctggtgcc tcacatcgga tacggattcc
     4981 accagtacct gccttatcct gacggaatga gccctttcca ggccgccatg gtggacggat
     5041 ctggatacca ggtgcacaga accatgcagt tcgaggacgg cgctagcctg accgtgaact
     5101 acagatacac ctacgagggc agccacatca aaggcgaggc ccaagtgaaa ggcactggat
     5161 tccctgctga cggacctgtg atgaccaaca gcctgacagc tgccgactgg tgcagaagca
     5221 agaaaacata cccaaacgac aagaccatca tcagcacctt caagtggtct tacaccaccg
     5281 gaaacggaaa gagatacaga agcaccgcca gaaccaccta caccttcgcc aagcctatgg
     5341 ccgccaacta cctgaagaac cagcctatgt acgtgttcag aaagaccgag ctgaagcaca
     5401 gcaaaacaga gctgaacttc aaagagtggc agaaagcctt caccgacgtg atgggaatgg
     5461 acgagctgta caagtgaggc ctctcgagcc tctagaacta tagtgagtcg tattacgtag
     5521 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     5581 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     5641 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     5701 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     5761 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     5821 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     5881 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5941 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6001 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6061 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6841 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7261 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7921 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     7981 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     8041 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     8101 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     8161 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     8221 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     8281 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     8341 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     8401 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     8461 cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat
     8521 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     8581 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     8641 gccaattcaa tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg
     8701 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     8761 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     8821 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     8881 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     8941 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     9001 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     9061 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     9121 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     9181 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     9241 ctattaatag taacttggca agtacattac tattggaagt acgccagggt acattggcag
     9301 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     9361 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     9421 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg ccattctgcc
     9481 tggggacgtc ggagcaagct tg
//
