LOCUS       sequence-348530-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of Patiria miniata Rab7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  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 5457)
  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: pZS218
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..1039
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1074..1092
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1074..1091
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1139..1846
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1284..1302)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1552..1571
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2548..2682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2572..2591)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2626..2645
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2795..2815)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2795..2813)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2834..2850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2834..2850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2847..2869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2858..2874
                     /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(2882..2888,2889..2906,2907..2912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2927..2948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3065..3082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3236..3824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3316..3335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3995..4786,4787..4855))
                     /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     4618..4637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4856..4960)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4986..5441)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5123..5144)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5335..5354
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
       61 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
      121 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
      181 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
      241 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
      301 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
      361 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
      421 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
      481 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
      541 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
      601 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
      661 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
      721 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
      781 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
      841 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
      901 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
      961 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     1021 caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg
     1081 tgacactata gaatacaagc tacttgttct ttttgcagga tccactagtc atcgattgat
     1141 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1201 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1261 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1321 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     1381 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     1441 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1501 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     1561 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     1621 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     1681 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     1741 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     1801 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggaat tcggagccgg
     1861 cgcgccaatg tcttcaagga agaaggtttt attgaaggtc atcattctag gagacagtgg
     1921 tgtcggcaag acctcgttga tgaaccaata tgtgaacaag aagtttagta accagtacaa
     1981 ggccaccatc ggcgctgact tcctgacaaa agaggtcatg gtagacgaca gactggtcac
     2041 aatgcagata tgggacacag ctgggcagga gaggttccag agtctgggtg tagcgttcta
     2101 ccgcggcgca gactgctgcg tcctcgtctt tgacgtgacc aatcagaatt ccttcaagtc
     2161 cctagacagc tggcgtgatg agtttctgat ccaagccagc cccagagatc ctgagaattt
     2221 ccccttcgtc gtactaggca acaagataga catggagaat agacaagttt cgaccaaacg
     2281 ggcgcagaac tggtgtggca gcaagaacaa catcccttac tttgagacca gcgctaagga
     2341 gagtatcaac gtggagcagg ccttccagac gatagccaag aacgccctgg cccaggaacc
     2401 cgagacagac atgtacaatg atttccccag ccagatcaaa ctcaccgatg acaccaaaca
     2461 ggataaatca gggtgttcat gttaaggccg gccagcgatc gccaaggcct ctcgagcctc
     2521 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     2581 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2641 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2701 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     2761 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     2821 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2881 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2941 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3001 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3061 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3121 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3181 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3241 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3301 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3361 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3421 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3481 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3541 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3601 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3661 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3721 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3781 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3841 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3901 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3961 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4021 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4081 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4141 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4201 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4261 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4321 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4381 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4441 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4501 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4561 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4621 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4681 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4741 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4801 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4861 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4921 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4981 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5041 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5101 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5161 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5221 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5281 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5341 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5401 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcag
//
