LOCUS       sequence-292665-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dinoflagelate backbone for gfp expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  AUTHORS   Sprecher BN, Zhang H, Lin S
  TITLE     Nuclear Gene Transformation in the Dinoflagellate Oxyrrhis marina.
  JOURNAL   Microorganisms. 2020 Jan 16;8(1). pii: microorganisms8010126. doi: 
            10.3390/microorganisms8010126.
  PUBMED    31963386
REFERENCE   2  (bases 1 to 5855)
  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: 
            "Microorganisms."; date: "2020-01-16"; volume: "8(1). pii: 
            microorganisms8010126. doi"; pages: " 10.3390/microorganisms8010126"
COMMENT     Sequence Label: DinoIII-gfp
FEATURES             Location/Qualifiers
     source          1..5855
                     /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             join(3919..3921,3922..3924,3925..4638)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3952..3980)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4575..4595
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5461..5478)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5461..5477)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5470..5492)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5686..5705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5805..5827
                     /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 ggctagcaaa ggagaagaac ttttcactgg agttgtccca
     3961 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     4021 gaaggtgatg ctacatacgg aaagcttacc cttaaattta tttgcactac tggaaaacta
     4081 cctgttccat ggccaacact tgtcactact ttctcttatg gtgttcaatg cttttcccgt
     4141 tatccggatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     4201 caggaacgca ctatatcttt caaagatgac gggaactaca agacgcgtgc tgaagtcaag
     4261 tttgaaggtg atacccttgt taatcgtatc gagttaaaag gtattgattt taaagaagat
     4321 ggaaacattc tcggacacaa actcgagtac aactataact cacacaatgt atacatcacg
     4381 gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cattgaagat
     4441 ggatccgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     4501 cttttaccag acaaccatta cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa
     4561 aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg
     4621 gatgagctct acaaatgatg atctcggcca ggagtcacag aaaacaagat cacttggaga
     4681 tgtttcaatc ccgacttgtg tcgtgccaga gtgctacttg aaaacttgaa aattgcggac
     4741 tgtcatggat tcgcccttgt cttgtgatcc tttttttggg ggagccaggt gagaacaatg
     4801 ttgtcgatgt gcttatttgg cttcgcagtc aaaacatggg atacttgaga catgaaagaa
     4861 aaatgccgca acgatagctc catccaattc cattcagctc cgactacaga tgatagcgct
     4921 tgacaccaat gacatgcttg tacagctgcc atttggaagg cagggaagct ccataagctc
     4981 gggtccccag gactttggtc ggtctcacat cagattcggc tagccagccc atagcagccg
     5041 cgggagattt cggttgtttg ctacaatgat tggggcgcct ttctgcgaac tttgtgacat
     5101 gtttcctcaa aatgtcaagc aattttgatc ttaaaagttt tgataatgct tgcttccaca
     5161 agcgacctac agtaggaaat gtctccacaa tctccacaga ttcaggactc atcactatgt
     5221 gtgccgtgca ggggtagggc gcagacatga caacatacaa cacacatgaa ctaaagaatc
     5281 caagtcgcgg acaaaaaaat ctgatcttac acttacacag aatgcaggtt attagcgacg
     5341 cttccattgc caccggagtg gcaatcgttg aggcgcttca tcgaacagag ggtgaacttc
     5401 ttgaggctgg gaggaccgcg cagatgcggc tgataaaagt ttcacaagca cacggaattc
     5461 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5521 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5581 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     5641 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     5701 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5821 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
