LOCUS       sequence-348527-        8183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of Patiria miniata Lamp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8183)
  AUTHORS   Swartz SZ, Tan TH, Perillo M, Fakhri N, Wessel GM, Wikramanayake AH,
            Cheeseman IM
  TITLE     Polarized Dishevelled dissolution and reassembly drives embryonic 
            axis specification in sea star oocytes.
  JOURNAL   Curr Biol. 2021 Dec 20;31(24):5633-5641.e4. doi: 
            10.1016/j.cub.2021.10.022. Epub 2021 Nov 4.
  PUBMED    34739818
REFERENCE   2  (bases 1 to 8183)
  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: 
            "10.1016/j.cub.2021.10.022"; journalName: "Curr Biol"; date: 
            "2021-12-20- 20"; volume: "31"; issue: "24"; pages: "5633-5641.e4"
COMMENT     Sequence Label: pZS222
FEATURES             Location/Qualifiers
     source          1..8183
                     /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"
     CDS             join(2214..2216,2217..2219,2220..2930)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKFAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2937..2939,2940..2942,2943..3653)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2937..2939,2940..2942,2943..3653)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3660..3662,3663..3665,3666..4379)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3660..3662,3663..3665,3666..4376)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4386..4520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4410..4429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4464..4483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4633..4653)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4633..4651)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4672..4688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4672..4688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4685..4707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4696..4712
                     /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(4720..4726,4727..4744,4745..4750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4765..4786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4903..4920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5074..5662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5154..5173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5833..6624,6625..6693))
                     /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     6456..6475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6694..6798)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6824..7279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6961..6982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7173..7192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccactagt ggcgcgccat gaagcttctt acagcagtcc
      121 tacttcttat ttttgtagcg cttggcgcag cgcagaacac cgagtctgca gccaccactg
      181 ctcctactaa gaaatccaca gagggaccca cttcaaaggc aaaaactgca gggccagcaa
      241 caacgggggc tacaaccaaa gcaccaaagg ttacaacagc aactgcagca actgcagtac
      301 cgacaacagc aacacctgca tcaactgcaa ctgcaactgc ggcaactgca gggccagcaa
      361 caactgctgg gactgccact gcgccaactg cagggccagc aacaaatgca agcaccgcag
      421 cacccaaaac gacggcagca cccacaagcc cgacaccggg ccccaccacc gtgccgcccc
      481 cattttttac ctataacgtg acgagcgccg atgggcaggt gtgctttctg gcccagtttt
      541 atgcagggtt ctcaggcacg tatgagctca agaacacatc catgatgaac tttaactttg
      601 agatgccaga gaacgccaaa gtcgatgcca agaagagcag gtgtggctgg ccttcacaaa
      661 ccgccatcat gttcctgacc tggaccgaag gtgcagacag cttggctctg gaattcagca
      721 tggacaacga agcccaaacc tttcagctga aatctgccaa cgctagtctg acgcattccc
      781 ctgcacactt ccctgatagc aaaactacag gggcagtcaa gtacggcggt gtcaccatgg
      841 gagacttctt caaggctgat tttggataca gcttcaagtg tgcgtccacc gagcagattg
      901 cttacaaggc caacgacttt aagtttacct tctcccagct gaaagtgcag ccgtatgaag
      961 tggacaaaac attcggccaa gagagccctt gcgcagcaga cggagcccac tggtccctga
     1021 aagagaaggg taacttttgc ctgctggcag atttccaact gaccttcacc ggggaataca
     1081 caaaaaacga ctcgaaaatg ggtacgttta ccgtcatgct ccccgcagat gcccgtatcg
     1141 acctggccaa cagtcattgt gaggatagca acgcccctat ggcccaggct gtcttggcac
     1201 tgaccttcga tcaaaactcc ttcatgcagg tgtaccaagt cgacaacaag acgaaaacct
     1261 tctcccttgt caacgccacg gctaagctga tgcacgataa aacccacttc ccggatagca
     1321 aggacgctgg taagatcgtt acttacggtg gcgcgcttac ggatcagttc cagactggtc
     1381 taggggatta ctacaagtgc tcggccatgc agaaggtaga attcaaggag aacaagtttg
     1441 ctctgagcta caccgacttg aagatccaac cgtttgatgt gcacatgacc ttcggaaact
     1501 tgagccagtg cggtcaggac acgccaactg ccaagccccc cacggtccct atgggtaact
     1561 ggactatcac aagagacaac aagacctgca tgatgatgca gtttcaagct cagttcatca
     1621 tctcctacca aaaggaagtc gggggtaaaa ctgagatgag caaagtgggt tactaccttc
     1681 ccaacaccac cctagacacc gccgagtcca cctgcggcga cacttcggcc atgtttgtcg
     1741 gcgccttctc ccctgaaaac ctgaagccca tcaaagactg ggtcatgacc ctggaattca
     1801 ccatgaagaa agacaagagc tactccatga caaacttcag cctggagttc atgatggatg
     1861 acgcctactt cccgggcctc aacaaatcaa tggccaagat gatgaccgtc ttcaacagca
     1921 cgacgcgctt tgattccaca gctggcaatt actacatgtg taacgcaaac caggacatca
     1981 tggccatgaa cgtcaacgca accttcatga agctcaagct gcagcccttc gctgagcagg
     2041 tcaagggcgg aggcctcgga agtccctctg actgtgctgc agacaatgac gtcagtaaca
     2101 tcgtgccgat cgccgtgggt tgcgccctgg caggcctggt catcatcgtc ctgattgcct
     2161 atctgatcgg gcgaagccgc tccaaccgat ccgggtacca gtcagtctct agaatggtga
     2221 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2281 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2341 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2401 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2461 acttcttcaa gttcgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2521 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2581 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2641 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     2701 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     2761 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2821 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2881 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag acgcgtatgg
     2941 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3001 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3061 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3121 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     3181 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3241 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3301 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3361 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3421 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3481 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3541 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3601 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagcccggga
     3661 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3721 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3781 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3841 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3901 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3961 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     4021 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     4081 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     4141 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     4201 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4261 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4321 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     4381 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4441 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4501 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4561 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     4621 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     4681 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat
     4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     6841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     7081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7261 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7321 tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg
     7381 actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg
     7441 actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg
     7501 caccatgcca attcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
     7561 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     7621 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     7681 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     7741 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     7801 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     7861 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     7921 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     7981 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     8041 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     8101 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     8161 taagcaatgc tcgtttaggg aac
//
