LOCUS       sequence-401164-        7102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7102)
  AUTHORS   Taelman VF, Dobrowolski R, Plouhinec JL, Fuentealba LC, Vorwald PP, 
            Gumper I, Sabatini DD, De Robertis EM
  TITLE     Wnt signaling requires sequestration of glycogen synthase kinase 3 
            inside multivesicular endosomes.
  JOURNAL   Cell. 2010 Dec 23. 143(7):1136-48.
  PUBMED    21183076
REFERENCE   2  (bases 1 to 7102)
  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: "Cell. 2010 
            Dec 23. 143(7):1136-48."
COMMENT     Sequence Label: pCS2 HRS-RFP
FEATURES             Location/Qualifiers
     source          1..7102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        527..1511
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1546..1564
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1546..1563
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1605..1631
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1759..1777)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2027..2046
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2221..2244
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4665..4799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4689..4708)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4743..4762
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4912..4932)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4912..4930)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4951..4967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4951..4967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4964..4986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4975..4991
                     /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(4999..5005,5006..5023,5024..5029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5044..5065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5182..5199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5433..5452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6112..6903,6904..6972))
                     /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     6735..6754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6973..7077)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
      541 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
      601 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
      661 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
      721 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
      781 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
      841 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
      901 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
      961 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     1021 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     1081 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     1141 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     1201 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     1261 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     1321 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     1381 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     1441 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     1501 gtttagggaa ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta
     1561 tagaatacaa gctacttgtt ctttttgcag gatccgccac catgtacccc tacgacgtcc
     1621 ccgactacgc agcctcctcc gaggacgtca tcaaggagtt catgcgcttc aaggtgcgca
     1681 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1741 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     1801 gggacatcct gtcccctcag ttccagtacg gctccaaggc ctacgtgaag caccccgccg
     1861 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1921 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1981 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     2041 agaagaccat gggctgggag gcctccaccg agcggatgta ccccgaggac ggcgccctga
     2101 agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac gccgaggtca
     2161 agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag accgacatca
     2221 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgcgccg
     2281 agggccgcca ctccaccgaa tcactagtga attcaaggcc tctcgagggg cgaggcagcg
     2341 gcaccttcga gcgtctccta gacaaggcga ccagccagct cctgttggag acagattggg
     2401 agtccatttt gcagatctgc gacctgatcc gccaagggga cacacaagca aaatatgctg
     2461 tgaattccat caagaagaaa gtcaacgaca agaacccaca cgtcgccttg tatgccctgg
     2521 aggtcatgga atctgtggta aagaactgtg gccagacagt tcatgatgag gtggccaaca
     2581 agcagaccat ggaggagctg aaggacctgc tgaagagaca agtggaggta aacgtccgta
     2641 acaagatcct gtacctgatc caggcctggg cgcatgcctt ccggaacgag cccaagtaca
     2701 aggtggtcca ggacacctac cagatcatga aggtggaggg gcacgtcttt ccagaattca
     2761 aagagagcga tgccatgttt gctgccgaga gagccccaga ctgggtggac gctgaggaat
     2821 gccaccgctg cagggtgcag ttcggggtga tgacccgtaa gcaccactgc cgggcgtgtg
     2881 ggcagatatt ctgtggaaag tgttcttcca agtactccac catccccaag tttggcatcg
     2941 agaaggaggt gcgcgtgtgt gagccctgct acgagcagct gaacaggaaa gcggagggaa
     3001 aggccacttc caccactgag ctgccccccg agtacctgac cagccccctg tctcagcagt
     3061 cccagctgcc ccccaagagg gacgagacgg ccctgcagga ggaggaggag ctgcagctgg
     3121 ccctggcgct gtcacagtca gaggcggagg agaaggagag gctgagacag aagtccacgt
     3181 acacttcgta ccccaaggcg gagcccatgc cctcggcctc ctcagcgccc cccgccagca
     3241 gcctgtactc ttcacctgtg aactcgtcgg cgcctctggc tgaggacatc gaccctgagc
     3301 tcgcacggta tctcaaccgg aactactggg agaagaagca ggaggaggct cgcaagagcc
     3361 ccacgccatc tgcgcccgtg cccctgacgg agccggctgc acagcctggg gaagggcacg
     3421 cagcccccac caacgtggtg gagaaccccc tcccggagac agactctcag cccattcctc
     3481 cctctggtgg cccctttagt gagccacagt tccacaatgg cgagtctgag gagagccacg
     3541 agcagttcct gaaggcgctg cagaacgccg tcaccacctt cgtgaaccgc atgaagagta
     3601 accacatgcg gggccgcagc atcaccaatg actcggccgt gctctcactc ttccagtcca
     3661 tcaacggcat gcacccgcag ctgctggagc tgctcaacca gctggacgag cgcaggctgt
     3721 actatgaggg gctgcaggac aagctggcac agatccgcga tgcccggggg gcgctgagtg
     3781 ccctgcgcga agagcaccgg gagaagcttc gccgggcagc cgaggaggca gagcgccagc
     3841 gccagatcca gctggcccag aagctggaga taatgcggca gaagaagcag gagtacctgg
     3901 aggtgcagag gcagctggcc atccagcgcc tgcaggagca ggagaaggag cggcagatgc
     3961 ggctggagca gcagaagcag acggtccaga tgcgcgcgca gatgcccgcc ttccccctgc
     4021 cctacgccca gctccaggcc atgcccgcag ccggaggtgt gctctaccag ccctcgggac
     4081 cagccagctt ccccagcacc ttcagccctg ccggctcggt ggagggctcc ccaatgcacg
     4141 gcgtgtacat gagccagccg gcccctgccg ctggccccta ccccagcatg cccagcactg
     4201 cggctgatcc cagcatggtg agtgcctaca tgtacccagc aggggccact ggggcgcagg
     4261 cggcccccca ggcccaggcc ggacccaccg ccagccccgc ttactcatcc taccagccta
     4321 ctcccacagc gggctaccag aacgtggcct cccaggcccc acagagcctc ccggccatct
     4381 ctcagcctcc gcagtccagc accatgggct acatggggag ccagtcagtc tccatgggct
     4441 accagcctta caacatgcag aatctcatga ccaccctccc aagccaggat gcgtctctgc
     4501 caccccagca gccctacatc gcggggcagc agcccatgta ccagcagatg gcaccctctg
     4561 gcggtccccc ccagcagcag ccccccgtgg cccagcaacc gcaggcacag gggccgccgg
     4621 cacagggcag cgaggcccag ctcatttcat tcgactgata cgtagatcca gacatgataa
     4681 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4741 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4801 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     4861 aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     4921 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4981 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     5041 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5101 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7081 atttccccga aaagtgccac ct
//
