LOCUS       sequence-307969-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dinoflagelate backbone for amino 3'-glycosyl phosphotransferase 
            (neo) gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  AUTHORS   Faktorova D, Nisbet RER, Fernandez Robledo JA, Casacuberta E, Sudek 
            L, Allen AE, Ares M Jr, Areste C, Balestreri C, Barbrook AC, 
            Beardslee P, Bender S, Booth DS, Bouget FY, Bowler C, Breglia SA, 
            Brownlee C, Burger G, Cerutti H, Cesaroni R, Chiurillo MA, Clemente 
            T, Coles DB, Collier JL, Cooney EC, Coyne K, Docampo R, Dupont CL, 
            Edgcomb V, Einarsson E, Elustondo PA, Federici F, Freire-Beneitez V,
            Freyria NJ, Fukuda K, Garcia PA, Girguis PR, Gomaa F, Gornik SG, Guo
            J, Hampl V, Hanawa Y, Haro-Contreras ER, Hehenberger E, Highfield A,
            Hirakawa Y, Hopes A, Howe CJ, Hu I, Ibanez J, Irwin NAT, Ishii Y, 
            Janowicz NE, Jones AC, Kachale A, Fujimura-Kamada K, Kaur B, Kaye 
            JZ, Kazana E, Keeling PJ, King N, Klobutcher LA, Lander N, Lassadi 
            I, Li Z, Lin S, Lozano JC, Luan F, Maruyama S, Matute T, Miceli C, 
            Minagawa J, Moosburner M, Najle SR, Nanjappa D, Nimmo IC, Noble L, 
            Novak Vanclova AMG, Nowacki M, Nunez I, Pain A, Piersanti A, 
            Pucciarelli S, Pyrih J, Rest JS, Rius M, Robertson D, Ruaud A, 
            Ruiz-Trillo I, Sigg MA, Silver PA, Slamovits CH, Jason Smith G, 
            Sprecher BN, Stern R, Swart EC, Tsaousis AD, Tsypin L, Turkewitz A, 
            Turnsek J, Valach M, Verge V, von Dassow P, von der Haar T, Waller 
            RF, Wang L, Wen X, Wheeler G, Woods A, Zhang H, Mock T, Worden AZ, 
            Lukes J
  TITLE     Genetic tool development in marine protists: emerging model 
            organisms for experimental cell biology.
  JOURNAL   Nat Methods. 2020 May;17(5):481-494. doi: 10.1038/s41592-020-0796-x.
            Epub 2020 Apr 6.
  PUBMED    32251396
REFERENCE   2  (bases 1 to 5930)
  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/s41592-020-0796-x"; journalName: "Nat Methods"; date: 
            "2020-05"; volume: "17"; issue: "5"; pages: "481-494"
COMMENT     Sequence Label: DinoIII-neo
FEATURES             Location/Qualifiers
     source          1..5930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3919..4713
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3919..4713
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3973..3992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4583..4602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5536..5553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5536..5552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5545..5567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5761..5780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5880..5902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgacga ttcccgtagc cattttggct
     2281 caaggtacaa gtcgggctga tgcggtcacg caggcctctt ttgtatcaga taagacagca
     2341 gacgtacatg atataaatat ttattatcgc gatgctccat ctgcaaaatt ccacttggca
     2401 gagaagattt tcaatgcatg aaaacctcgc agaaccaagc accaatgtca gatcatgtat
     2461 gagtcctagg cgccagtaag tctttcaatt gggtgatggc cacattacct gttctcttcc
     2521 tgagggatgc agaatgtggt gtgagtggtt gccgaatgct gtgctgaaca tgtagatggg
     2581 ctgccccagc caagtggata acctacaagg actgcaggat gtagatccag cagtcctggt
     2641 agataatgat tcagacaccg gctcaggctg gcaacagagc aggcaaacct gaacactatc
     2701 gcgcagtggt gggaatacag cccagaatgg gaagcaacct ggcctcttca gccagccatg
     2761 accacaagca actctgcagt tttgcaacac gaaatctttc agggtctgct ccctgtgctt
     2821 acacatattc aaagatattc ttctgcgcat ttcaatgcaa attcttggcg atgattcgct
     2881 tccgcaggta tagtgttact agtatataag tctaatcata tacttcaatt tattagtaat
     2941 attgtgaatt tctaggcaag aaggatgcct ggactctgaa ttattatggc attgagtaga
     3001 atctggatct tgatgattat gcattaatat cttgaaatgc atttggattc ccttcgggga
     3061 tcatccgtta aaattggaac gatacagaga agattagcat ggcccctgcg caaggatgac
     3121 acgcacaaat cgagaagtgt aaacaatttt tttgaaatta attgccactt tattttgaat
     3181 acctgaatat gcaggtgaag tagtataagg tattcattat cattgcgatt ttctagtaag
     3241 tcccgtggct gtcatatcta gtaaccctca cctggcaggt ggggaaaagg cgaagacaaa
     3301 tagaatcaaa tagaatgtat gtgctgacgt agggctacat tacctgaggc ttgaggaacc
     3361 tgtgggtatg aatgctactg ttggcagcat ccagtaggtt cagaaacaca cgtccccacg
     3421 taattttttg catgtctaaa tgcactgtga tgattgatga tgattgattc cttgaaggcc
     3481 atggtgggtt actctgtggc ctcttgccac tgacttccag gaaaacgcgg attttctgac
     3541 catcacaagg gaccactacg agaagtatgc agcgatctca gcagagttca tttactgacc
     3601 attctcagga agatcgctcg ggatcacttg ggtaatattg ctttcttcgc agatgttttt
     3661 tccctattaa gctttttgag tcccctcatc gagtgccgca taagtttttg cgttgacatt
     3721 ggcggcaaaa agtaagacta aacgatagtt gcttcaagca cgcttctcaa tcaacatttt
     3781 ttcaaatcaa atgggcacag gcaggtacca tccagtcccg cagaagtgct tatatatata
     3841 tatattatac actcacgtga gtgtgttcaa tgcaggtctg ttcctcaagt gcttagacca
     3901 aatttcggag cctctagtat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3961 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4021 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4081 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4141 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4201 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4261 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4321 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4381 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4441 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4501 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4561 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4621 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4681 gccttctatc gccttcttga cgagttcttc tgatgatctc ggccaggagt cacagaaaac
     4741 aagatcactt ggagatgttt caatcccgac ttgtgtcgtg ccagagtgct acttgaaaac
     4801 ttgaaaattg cggactgtca tggattcgcc cttgtcttgt gatccttttt ttgggggagc
     4861 caggtgagaa caatgttgtc gatgtgctta tttggcttcg cagtcaaaac atgggatact
     4921 tgagacatga aagaaaaatg ccgcaacgat agctccatcc aattccattc agctccgact
     4981 acagatgata gcgcttgaca ccaatgacat gcttgtacag ctgccatttg gaaggcaggg
     5041 aagctccata agctcgggtc cccaggactt tggtcggtct cacatcagat tcggctagcc
     5101 agcccatagc agccgcggga gatttcggtt gtttgctaca atgattgggg cgcctttctg
     5161 cgaactttgt gacatgtttc ctcaaaatgt caagcaattt tgatcttaaa agttttgata
     5221 atgcttgctt ccacaagcga cctacagtag gaaatgtctc cacaatctcc acagattcag
     5281 gactcatcac tatgtgtgcc gtgcaggggt agggcgcaga catgacaaca tacaacacac
     5341 atgaactaaa gaatccaagt cgcggacaaa aaaatctgat cttacactta cacagaatgc
     5401 aggttattag cgacgcttcc attgccaccg gagtggcaat cgttgaggcg cttcatcgaa
     5461 cagagggtga acttcttgag gctgggagga ccgcgcagat gcggctgata aaagtttcac
     5521 aagcacacgg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5581 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5641 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     5701 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5821 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5881 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
