LOCUS       sequence-348522-        6947 bp    DNA     circular SYN 15-APR-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 6947)
  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 6947)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pZS212
FEATURES             Location/Qualifiers
     source          1..6947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2458..3442
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3477..3495
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3477..3494
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             5680..6390
                     /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(5825..5843)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6093..6112
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    6441..6575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6465..6484)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6519..6538
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6688..6708)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6688..6706)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6727..6743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6727..6743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6740..6762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6751..6767
                     /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(6775..6781,6782..6799,6800..6805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6820..6841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc
     2461 gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac
     2521 ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac
     2581 cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca
     2641 ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc
     2701 cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct
     2761 atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg
     2821 gccaggttca atactatgta ttggccctat gccatatagt attccatata tgggttttcc
     2881 tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta
     2941 ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat
     3001 tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac
     3061 ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg
     3121 tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg
     3181 acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca
     3241 ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta
     3301 aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac
     3361 gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat
     3421 aagcaatgct cgtttaggga accgccattc tgcctgggga cgtcggagca agcttgattt
     3481 aggtgacact atagaataca agctacttgt tctttttgca ggatccacta gtggcgcgcc
     3541 atgaagcttc ttacagcagt cctacttctt atttttgtag cgcttggcgc agcgcagaac
     3601 accgagtctg cagccaccac tgctcctact aagaaatcca cagagggacc cacttcaaag
     3661 gcaaaaactg cagggccagc aacaacgggg gctacaacca aagcaccaaa ggttacaaca
     3721 gcaactgcag caactgcagt accgacaaca gcaacacctg catcaactgc aactgcaact
     3781 gcggcaactg cagggccagc aacaactgct gggactgcca ctgcgccaac tgcagggcca
     3841 gcaacaaatg caagcaccgc agcacccaaa acgacggcag cacccacaag cccgacaccg
     3901 ggccccacca ccgtgccgcc cccatttttt acctataacg tgacgagcgc cgatgggcag
     3961 gtgtgctttc tggcccagtt ttatgcaggg ttctcaggca cgtatgagct caagaacaca
     4021 tccatgatga actttaactt tgagatgcca gagaacgcca aagtcgatgc caagaagagc
     4081 aggtgtggct ggccttcaca aaccgccatc atgttcctga cctggaccga aggtgcagac
     4141 agcttggctc tggaattcag catggacaac gaagcccaaa cctttcagct gaaatctgcc
     4201 aacgctagtc tgacgcattc ccctgcacac ttccctgata gcaaaactac aggggcagtc
     4261 aagtacggcg gtgtcaccat gggagacttc ttcaaggctg attttggata cagcttcaag
     4321 tgtgcgtcca ccgagcagat tgcttacaag gccaacgact ttaagtttac cttctcccag
     4381 ctgaaagtgc agccgtatga agtggacaaa acattcggcc aagagagccc ttgcgcagca
     4441 gacggagccc actggtccct gaaagagaag ggtaactttt gcctgctggc agatttccaa
     4501 ctgaccttca ccggggaata cacaaaaaac gactcgaaaa tgggtacgtt taccgtcatg
     4561 ctccccgcag atgcccgtat cgacctggcc aacagtcatt gtgaggatag caacgcccct
     4621 atggcccagg ctgtcttggc actgaccttc gatcaaaact ccttcatgca ggtgtaccaa
     4681 gtcgacaaca agacgaaaac cttctccctt gtcaacgcca cggctaagct gatgcacgat
     4741 aaaacccact tcccggatag caaggacgct ggtaagatcg ttacttacgg tggcgcgctt
     4801 acggatcagt tccagactgg tctaggggat tactacaagt gctcggccat gcagaaggta
     4861 gaattcaagg agaacaagtt tgctctgagc tacaccgact tgaagatcca accgtttgat
     4921 gtgcacatga ccttcggaaa cttgagccag tgcggtcagg acacgccaac tgccaagccc
     4981 cccacggtcc ctatgggtaa ctggactatc acaagagaca acaagacctg catgatgatg
     5041 cagtttcaag ctcagttcat catctcctac caaaaggaag tcgggggtaa aactgagatg
     5101 agcaaagtgg gttactacct tcccaacacc accctagaca ccgccgagtc cacctgcggc
     5161 gacacttcgg ccatgtttgt cggcgccttc tcccctgaaa acctgaagcc catcaaagac
     5221 tgggtcatga ccctggaatt caccatgaag aaagacaaga gctactccat gacaaacttc
     5281 agcctggagt tcatgatgga tgacgcctac ttcccgggcc tcaacaaatc aatggccaag
     5341 atgatgaccg tcttcaacag cacgacgcgc tttgattcca cagctggcaa ttactacatg
     5401 tgtaacgcaa accaggacat catggccatg aacgtcaacg caaccttcat gaagctcaag
     5461 ctgcagccct tcgctgagca ggtcaagggc ggaggcctcg gaagtccctc tgactgtgct
     5521 gcagacaatg acgtcagtaa catcgtgccg atcgccgtgg gttgcgccct ggcaggcctg
     5581 gtcatcatcg tcctgattgc ctatctgatc gggcgaagcc gctccaaccg atccgggtac
     5641 cagtcagtcg gccggccagc gatcgccgga catcgattga tggtgagcaa gggcgaggag
     5701 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     5761 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     5821 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     5881 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     5941 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     6001 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     6061 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     6121 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     6181 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     6241 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     6301 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     6361 accggcggca tggacgagct gtacaagtaa gaattcaagg cctctcgagc ctctagaact
     6421 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
     6481 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     6541 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     6601 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc
     6661 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     6721 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6781 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6901 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgct
//
