LOCUS       sequence-80602-e        5884 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 5884)
  AUTHORS   Levskaya A, Weiner OD, Lim WA, Voigt CA
  TITLE     Spatiotemporal control of cell signalling using a light-switchable 
            protein interaction.
  JOURNAL   Nature. 2009 Oct 15;461(7266):997-1001.
  PUBMED    19749742
REFERENCE   2  (bases 1 to 5884)
  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: "Nature."; 
            date: "2009-10-15"; volume: "461(7266)"; pages: "997-1001"
COMMENT     Sequence Label: pAL197
FEATURES             Location/Qualifiers
     source          1..5884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        169..379
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      238..373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    complement(660..684)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             698..1408
                     /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(843..861)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1111..1130
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(1436..1454)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1437..1454)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1474..1491)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1480..1704
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1750..2178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1837..1856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2047..2068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2187..2207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2192..2522
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2373..2508
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2589..3383
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2643..2662)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3253..3272
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3557..3678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3594..3613)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3648..3667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3727..3743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3727..3743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3740..3762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3751..3767
                     /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(3775..3781,3782..3799,3800..3805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3820..3841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3958..3975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4129..4717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4209..4228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4888..5679,5680..5748))
                     /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     5511..5530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5749..5853)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgaggc aggcagaagt
      181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
      361 ttttttggag gcctaggcta agcttggtac catgaagaaa gagaaagagc ggccagagat
      421 ttctctccct tcagattttg aacacacaat tcatgtcggt tttgatgctg tcacagggga
      481 gtttacggga atgccagagc agtgggcccg cttgcttcag acatcaaata tcactaagtc
      541 ggagcagaag aaaaacccgc aggctgttct ggatgtgttg gagttttaca actcgaagaa
      601 gacatccaac agccagaaat acatgagctt tacagataag tcaaagggtg ggcgcgccga
      661 cccagctttc ttgtacaaag tggtgagctc agccaccatg gtgagcaagg gcgaggagga
      721 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      781 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      841 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      901 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
      961 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     1021 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     1081 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     1141 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     1201 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     1261 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     1321 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     1381 cggcggcatg gacgagctgt acaagtaact cgagcatgca tctagagggc cctattctat
     1441 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     1501 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1561 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1621 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1681 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     1741 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1801 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1861 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     1921 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1981 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2041 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2101 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     2161 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2221 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2281 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2341 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2401 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2461 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2521 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     2581 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2641 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2701 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2761 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2821 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2881 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     2941 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3001 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3061 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3121 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3181 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3241 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3301 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3361 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3421 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3481 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3541 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3601 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3661 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     3721 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3781 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3901 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3961 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4021 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4081 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4141 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4201 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4261 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4321 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4381 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4441 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4501 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4561 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4621 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4681 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4741 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4801 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4861 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4921 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4981 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5041 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5101 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5161 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5221 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5281 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5341 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5401 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5461 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5521 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5581 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5641 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5701 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5761 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5821 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5881 cgtc
//
