LOCUS       sequence-482908-        6487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AmNeonGreen-13myc in Exaiptasia diaphana; In vitro 
            transcription of the same construct via SP6 polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6487)
  AUTHORS   n/a
  TITLE     Aiptasia Genetic Tools Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6487)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCR241
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(64..75,76..238)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     promoter        2449..2467
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2449..2466
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3203..3700
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     polyA_signal    3760..3894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3784..3803)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3838..3857
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4146..4734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4226..4245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4905..5696,5697..5765))
                     /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     5528..5547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5766..5870)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(6197..6384,6385..6396)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6432..6452)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6432..6450)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6471..6487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6471..6487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgttcagac acccatatcc actctgttcc acacaggtca gaggtttgtc caggagttct
       61 tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa gtacttaggg
      121 aaaattttac tcaattaaaa gtaaaagtat ctggctagaa tcttacttga gtaaaagtaa
      181 aaaagtactc cattaaaatt gtacttgagt attaaggaag taaaagtaaa agcaagaaag
      241 aaaactagag attcttgttt aagcttttaa tctcaaaaaa cattaaatga aatgcataca
      301 aggttttatc ctgctttaga actgtttgta tttaattatc aaactataag acagacaatc
      361 taatgccagt acacgctact caaagttgta aaacctcaga tttaacttca gtagaagctg
      421 attctcaaaa ttgttagtgt caagcctagc tcttttgggg ctgaaaagca atcctgcagt
      481 gctgaaaagc ctctcacagg cagccgatgc gggaagaggt gtattagtct tgatagagag
      541 gctgcaaata gcaggaaacg tgagcagaga ctccctggtg tctgaaacac aggccagata
      601 ggccccttac gtacgctagc gatatcggat ccgtcgactg cattcactgt tcttaatatt
      661 gtataatagg caaaaatcga ggtcacagat ctcgaagaaa attggtaaaa taaacaaaat
      721 agagaataat taatctaatg aatcgaaaaa ctcgttttca gacattcaaa taagaaacaa
      781 aatgtcacat aatggatata aacaagtcaa tcacaatgca aaacctccat caccggaggc
      841 tccggtcgga ttgtgtgaga tgggaagggt acaaaggctc atgggtgtct atagcaagtt
      901 tacacaggga tcccaaatat atttatactc ttttaactgt ttggatttat tattcttgac
      961 tttcacatga cgtcatcagt caaaaaaaaa atctaaaatc acagacacac aacattgtgg
     1021 tactgaaata tcttgaagag catgatagaa gttcgctctg aatgtttaaa ctttttttgc
     1081 tgtttattca ttgagttatt agcgttcaaa aatatgttac gtaatactgc cacgcgactt
     1141 gatacacgcc aaaaatactc gaaaagccta tgcatcttaa aacaattata gtttcagcat
     1201 caatttaaat gttaacaatg atattgttgt aaagttaaag caaaatcatc agttttgagc
     1261 tcaattcgat acggtccgcc attttgaagc ccctgacggg gacacaatca tggcggcccc
     1321 atacatttct ttgtaaagta agaccgtatt caaacgctta caattttgct aggaaatcac
     1381 agttttgatt caaactttac ataactgttg ggagttatat acttttttga tccctgaact
     1441 taaaaaatta atttgaatct taaaacaaag ctttttgtag tcatttcttg atcagtccaa
     1501 aacgcattga gccgtgctaa tcccatacat tcttaccaat tttcgatcgc tcgtatttct
     1561 gttaaaattt attatttcgg ctcaaaattt cgtcatgatc cttaaattaa gatgttctct
     1621 cgagagcatt aataaaagtt tgctaattct gcctctaaat aggtttttta gactatcacg
     1681 tgatcagtga aagtcaagaa ttgattggct atgaaattat aagaaactat gatatttatt
     1741 agttttttgg taagctgaaa ttctaaagca atcaaaatat ttttcttgtc tgccagcgcc
     1801 atttttttcg ataagtcact ttgtcgaaga aagaagaaac acctaagata gaatctaaaa
     1861 tcaaaagcaa aaactactag agctatgcta aagccaagaa gtttgcaaaa aaacttgtga
     1921 atattaaaat tattgaaatt gaaattaact tcggcctttt tcaggccttt gaaaacgcaa
     1981 acaaaagcca gtaaatgaac cagaaaagac gccaagaaac taactttgtt tgctatggtc
     2041 attattctta attgaaaaca tatgaaatca caatttagca tactttttgt ctacttttgt
     2101 tcggagtctg gacagcgtgc tctgtcacgc tgtaagctgt gactcttttg ttacatctcc
     2161 attgttgaac tgtcaagtgt ttcgtagcaa cccaagtctt caatttgtga ttggtttatt
     2221 cattcttccg tagcgatatt taagacaacg cattcttaat caccgcctct tttacatcat
     2281 ccaaggccac gtgtgcaact tgtgagtttt tctcttactt ttactgccct tttctcccct
     2341 ttttaactca aattatcgtt acttaagcga attgtgacat tcgttgaact gttttcttta
     2401 aaatttgtat tttagatcac tgttgaatcg ctcggcaact tttcaaatat ttaggtgaca
     2461 ctatagaagc gctggtacca taatggctat ggtgagcaag ggcgaggagg ataacatggc
     2521 ctctctccca gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga
     2581 catggtgggt cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctaaagtc
     2641 caccaagggt gacctccagt tctccccctg gattctcgtc cctcatatcg ggtatggctt
     2701 ccatcagtac ctaccctacc ctgacgggat gagtcctttc caggccgcta tggtagatgg
     2761 ctccggatac caagtccatc gtacaatgca gtttgaagat ggtgcctccc ttactgttaa
     2821 ctacagatac acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg
     2881 tttccctgct gacggtcctg tgatgaccaa cagtctcacc gctgcggact ggtgcaggag
     2941 taagaagact taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac
     3001 tggaaatggc aagagatacc gtagcactgc gaggaccacc tacacctttg ctaagccaat
     3061 ggcggctaac tatctcaaga accagcctat gtacgtgttc cgtaagacgg aactcaagca
     3121 ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat
     3181 ggacgagctg tacaagggct ccgaacaaaa gctaatctcc gaggaagact tgaacggtga
     3241 acaaaaatta atctcagaag aagacttgaa cggactcgac ggtgaacaaa agttgatttc
     3301 tgaagaagat ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca
     3361 aaaattaatc tcagaagaag acttgaacgg actcgacggt gaacaaaagt tgatttctga
     3421 agaagatttg aacggtgaac aaaagctaat ctccgaggaa gacttgaacg gtgaacaaaa
     3481 attaatctca gaagaagact tgaacggact cgacggtgaa caaaagttga tttctgaaga
     3541 agatttgaac ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg aacaaaaatt
     3601 aatctcagaa gaagacttga acggactcga cggtgaacaa aagttgattt ctgaagaaga
     3661 tttgaacggt gaacaaaagc taatctccga ggaagacttg aacggtgaac aaaaattaat
     3721 ctagttgccc atggcatatg gtatacatcg atgagctgcg atccagacat gataagatac
     3781 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3841 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3901 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3961 aagtaaaacc tctacaaatg tggtatggct gattatgatc cgcggacgtg gccatctaga
     4021 attcagatct gacctgaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4081 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4141 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4201 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4261 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4321 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4381 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4441 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4501 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4561 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4621 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4681 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4741 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4801 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4861 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4981 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5041 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5101 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5341 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5401 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5581 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5761 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccg
     5881 cgaagatacg gccacgggtg ctcttgatcc tgtggctgat tttggactgt gctgctcgca
     5941 gctgctgatg aatcacatac ttcctccatt ttcttccact gattgactgt tataatttcc
     6001 ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga cgtcacttcc
     6061 aaaggaccaa tgaacatgtc tgaccaattt catataatgt gaaaacgatt ttcataggca
     6121 gaataaataa catttaaatt aaactgggca tcagcgcaat tcaattggtt tggtaatagc
     6181 aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat
     6241 tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta
     6301 ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt
     6361 aatcaagtaa aattactcaa gtactttaca cctctgcggc cgccaccgcg gtggagctcc
     6421 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     6481 tttcctg
//
