LOCUS       sequence-252732-        7451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  c-di-GMP-II switch cdGII-CGUAA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7451)
  AUTHORS   Xiang JS, Kaplan M, Dykstra P, Hinks M, McKeague M, Smolke CD
  TITLE     Massively parallel RNA device engineering in mammalian cells with 
            RNA-Seq.
  JOURNAL   Nat Commun. 2019 Sep 23;10(1):4327. doi: 10.1038/s41467-019-12334-y.
  PUBMED    31548547
REFERENCE   2  (bases 1 to 7451)
  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 
            Commun."; date: "2019-09-23"; volume: "10(1):4327. doi"; pages: " 
            10.1038/s41467-019-12334-y"
COMMENT     Sequence Label: pCS4110
FEATURES             Location/Qualifiers
     source          1..7451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..259
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             272..540
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             570..1271
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1328..1347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1372..1419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(1632..1651)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1823..2202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2203..2406
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2357..2377
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2451..2470
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2451..2469
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2500..2509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2506..3216
                     /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     2919..2938
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3403..3420)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3409..3633
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3766..3785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3917..3964
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3972..4994
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3972..4994
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5124..5245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5161..5180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5215..5234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5294..5310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5294..5310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5307..5329)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5318..5334
                     /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(5342..5348,5349..5366,5367..5372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5387..5408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5525..5542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5696..6284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5776..5795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6455..7246,7247..7315))
                     /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     7078..7097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7316..7420)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctgga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
       61 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
      121 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
      181 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
      241 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
      301 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
      361 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
      421 cctctgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
      481 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
      541 caagctgtga ccctcgagta ttcgccacca tgagcgagct gattaaggag aacatgcaca
      601 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
      661 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
      721 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
      781 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
      841 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
      901 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
      961 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     1021 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     1081 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     1141 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     1201 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     1261 agcttaattg atctgcatga gacgatctca tgctggagtt cttcgcccac cctaggggga
     1321 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     1381 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     1441 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     1501 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     1561 tcggggcggc aggccctgcc atagcctcag gttactcata gatctcccga tcccctatgg
     1621 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     1681 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     1741 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     1801 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     1861 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1921 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1981 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2041 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2101 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2161 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2221 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2281 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2341 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2401 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     2461 cactataggg agacccaagc tggctagcac gctacggtcg ccaccatggt gagcaagggc
     2521 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaagtgca catggagggc
     2581 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2641 acccagaccg ccaagctgaa agtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2701 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2761 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2821 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2881 aaagtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2941 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3001 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaagt caagaccacc
     3061 tacaaggcca agaagcccgt gcagctgccc ggagcctaca acgtcaacat caagttggac
     3121 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3181 cactccaccg gcggcatgga cgagctgtac aagtaactga tcataaatat agggcccgga
     3241 tccaaacaaa caaagctgtc accgagaaac tgtgaagtat atcttaaacc tgggcactta
     3301 aaagatatat ggagttagta gtgcaacctg ctcggtctga tgagtcccgt aaggacgaaa
     3361 cagcaaaaag aaaaataaaa aggtaccaaa cccgctgatc agcctcgact gtgccttcta
     3421 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3481 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3541 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3601 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3661 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3721 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3781 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3841 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3901 atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa
     3961 cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     4021 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     4081 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     4141 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     4201 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     4261 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     4321 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     4381 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     4441 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     4501 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     4561 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     4621 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     4681 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     4741 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     4801 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     4861 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     4921 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     4981 gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     5041 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5101 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5161 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5221 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5281 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5341 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5401 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5461 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5521 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5581 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5641 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5881 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6121 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7441 cacctgacgt c
//
