LOCUS       sequence-173314-        5271 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 5271)
  AUTHORS   Park TJ, Mitchell BJ, Abitua PB, Kintner C, Wallingford JB
  TITLE     Dishevelled controls apical docking and planar polarization of basal
            bodies in ciliated epithelial cells.
  JOURNAL   Nat Genet. 2008 Jul . 40(7):871-9.
  PUBMED    18552847
REFERENCE   2  (bases 1 to 5271)
  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: "Nat Genet. 
            2008 Jul . 40(7):871-9."
COMMENT     Sequence Label: pCS2+ centrin-RFP
FEATURES             Location/Qualifiers
     source          1..5271
                     /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             619..1293
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             619..1293
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(749..767)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1017..1036
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1211..1234
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1323..1457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1347..1366)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1401..1420
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1570..1590)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1570..1588)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1609..1625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1609..1625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1622..1644)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1633..1649
                     /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(1657..1663,1664..1681,1682..1687))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1702..1723
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1840..1857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2011..2599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2091..2110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2770..3561,3562..3630))
                     /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     3393..3412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3631..3735)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3761..4216)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3898..3919)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4110..4129
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4287..5271
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca tggcctctgt tctgcgtaaa
      121 ccagggttgg gtgcaactca gcgtaggaga acaggagcaa aacctgacct gacagaggaa
      181 caaaaaaagg agatccgtga agcatttgat ttgtttgata ctgatggaac tgggacaatt
      241 gatgtcaagg aactaaaggt ggctatgcgg gctttgggat ttgagccaaa aaaagaggaa
      301 atgaaaaaaa ttatatctga cattgacaaa gatggttctg gaattattga ctttgaggac
      361 tttctgtctc tcatgactca aaaaatgagt gaaaaagact caaaagagga aattatgaag
      421 gcttttcgtc tatttgatga tgacaatact ggcaaaatct ctttcaaaaa tctgaaaagg
      481 gtagccaagg aattgggaga aaaccttaca gatgaagagc tacaggaaat gattgatgaa
      541 gccgatcgcg atggagatgg tgaaatcaat gaacaagagt ttctcagaat tatgagaaag
      601 acaagccttt gctctagaat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc
      661 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      721 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
      781 cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc ctacgtgaag
      841 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      901 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      961 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1021 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta ccccgaggac
     1081 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac
     1141 gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag
     1201 accgacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1261 gagcgcgccg agggccgcca ctccaccggc gcctctagaa ctatagtgag tcgtattacg
     1321 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1381 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1441 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1501 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     1561 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     1621 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     1681 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1741 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1801 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1861 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1921 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1981 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2041 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2101 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2161 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2221 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2281 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2341 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2401 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2461 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2521 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2581 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2641 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2701 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2761 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2821 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2881 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2941 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3001 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3061 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3121 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3181 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3241 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3301 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3361 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3421 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3481 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3541 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3601 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3661 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3721 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     3781 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3841 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3901 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3961 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4021 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4081 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4141 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4201 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4261 gtgcgggcct cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact
     4321 catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact
     4381 cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac
     4441 catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca
     4501 cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca
     4561 tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc
     4621 aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg
     4681 ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg
     4741 gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt
     4801 caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat
     4861 atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt
     4921 gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc
     4981 cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa
     5041 tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg
     5101 cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc
     5161 cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc
     5221 ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
