LOCUS       sequence-382135-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Arl13b with mCardinal in mammalian cells and zebrafish 
            embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  AUTHORS   Aljiboury AA, Ingram E, Krishnan N, Ononiwu F, Pal D, Manikas J, 
            Taveras C, Hall NA, Da Silva J, Freshour J, Hehnly H
  TITLE     Rab8, Rab11, and Rab35 coordinate lumen and cilia formation during 
            zebrafish left-right organizer development.
  JOURNAL   PLoS Genet. 2023 May 15;19(5):e1010765. doi: 
            10.1371/journal.pgen.1010765. eCollection 2023 May.
  PUBMED    37186603
REFERENCE   2  (bases 1 to 6082)
  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 
            Genet."; date: "2023-05-15"; volume: "19(5):e1010765. doi"; pages: "
            10.1371/journal.pgen.1010765. eCollection 2023 May"
COMMENT     Sequence Label: pCS2-Arl13b-mCardinal
FEATURES             Location/Qualifiers
     source          1..6082
                     /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"
     regulatory      74..83
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1364..2098
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2134..2268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2158..2177)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2212..2231
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2381..2401)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2420..2436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2420..2436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2433..2455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2444..2460
                     /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(2468..2474,2475..2492,2493..2498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2513..2534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2651..2668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2822..3410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2902..2921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3581..4372,4373..4441))
                     /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     4204..4223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4442..4546)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4572..5027)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4709..4730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4921..4940
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5098..6082
                     /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 tttgccacca tggtgttcag tctgatggcc aactgctgca acttgttcaa
      121 gcggtggcgg gagcctgtca gaaaggtgac acttgtgatg gtcggccttg ataatgctgg
      181 taaaacagcc acagcaaagg gaatccaagg agagcatcct gaagacgtag ctcccactgt
      241 tggcttttct aaaattgatc tgagacaagg aaagttccaa gttaccatct ttgacttagg
      301 aggtggaaaa agaattcggg gaatttggaa gaattattat gctgaatcct atggggtaat
      361 atttgttgtg gattccagtg atgaggagag aatggaagaa acaaaggaga caatgtccga
      421 agtgctaaga cacccgagga tatcaggaaa gcctatattg gtgctggcca acaagcaaga
      481 caaggagggg gctttaggcg aagcggatgt gattgagtgt ctctcgctgg agaagctggt
      541 caacgagcac aagtgcctgt gtcagatcga accttgttca gcagtcttgg gatatggaaa
      601 gaaaattgac aagtccatta aaaaggggct ttattggcta ctgcatatca ttgcaaagga
      661 ctttgatgcc ttaagtgaac gcatccagaa agacacaact gaacagcgag ctcttgagga
      721 acaagagaaa cgtgagaggg ctgaacgagt ccggaagtta agggaagaaa gagaacgaga
      781 gcagacggaa cttgatggga ccagtggtct ggctgagatt gactcaggac cagtccttgc
      841 gaatcctttc caacccattg cagctgtaat cattgagaat gaaaagaagc aggaaaaaga
      901 gaaaaagaag cagaccgtgg agaaggacag tgatgttggt ctcctagaac ataaagtgga
      961 gccagagcaa gcagcgcctc agagtgaggc agattgctgc ctccaaaacc ctgacgagcg
     1021 agtggtggac agttacaggg aggcactgtc gcagcagctg gacagtgagg atgagcagga
     1081 ccagcgaggg tcagagtccg gggaaaacag taagaagaaa accaagaaac taagaatgaa
     1141 aaggagtcat cgggtagaac cagtcaatac agacgagtct actccaaaga gtcccacgcc
     1201 tccccaacct ccccctcctg ttggctgggg aacccccaaa gtcactagac ttccaaaact
     1261 tgagcctctt ggtgaaacac gtcacaatga tttctatggg aagccgctgc ctcccctggc
     1321 tgtgcgacag agacctaacg gtgatgctca ggacacgatc tcaatggtga gcaagggcga
     1381 ggagctgatc aaggagaaca tgcacatgaa gctgtacatg gaaggcaccg tgaacaacca
     1441 ccacttcaag tgcaccaccg aaggggaggg caagccctac gagggcaccc agacccagag
     1501 gattaaggtg gtggagggag gccccctgcc gttcgcattc gacatcctgg ccacctgctt
     1561 tatgtacggg agcaagacct tcatcaacca cacccagggc atccccgatt tctttaagca
     1621 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     1681 taccgttacc caggacacca gcctccagga cggctgcttg atctacaacg tcaagctcag
     1741 aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     1801 caccaccgag accctgtacc ccgctgacgg cggcctggaa ggcagatgcg acatggccct
     1861 gaagctcgtg ggcgggggcc acctgcactg caacctgaag accacataca gatccaagaa
     1921 acccgctaag aacctcaaga tgcccggcgt ctactttgtg gaccgcagac tggaaagaat
     1981 caaggaggcc gacaatgaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg
     2041 cgacctccct agcaaactgg ggcacaaact taatggcatg gacgagctgt acaagtaact
     2101 gcagtctaga actatagtga gtcgtattac gtagatccag acatgataag atacattgat
     2161 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2221 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2281 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc
     2341 gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg
     2401 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2461 ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga
     2521 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2581 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4561 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4621 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4681 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4741 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4801 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4861 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4921 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4981 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     5041 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc
     5101 gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac
     5161 ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac
     5221 cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca
     5281 ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc
     5341 cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct
     5401 atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg
     5461 gccaggttca atactatgta ttggccctat gccatatagt attccatata tgggttttcc
     5521 tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta
     5581 ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat
     5641 tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac
     5701 ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg
     5761 tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg
     5821 acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca
     5881 ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta
     5941 aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac
     6001 gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat
     6061 aagcaatgct cgtttaggga ac
//
