LOCUS       sequence-250222-        6584 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 6584)
  AUTHORS   Chisari M, Saini DK, Cho JH, Kalyanaraman V, Gautam N
  TITLE     G protein subunit dissociation and translocation regulate cellular 
            response to receptor stimulation.
  JOURNAL   PLoS One. 2009 Nov 11;4(11):e7797.
  PUBMED    19936219
REFERENCE   2  (bases 1 to 6584)
  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: "PLoS One.";
            date: "2009-11-11"; volume: "4(11)"; pages: "e7797"
COMMENT     Sequence Label: mcherry-PH
FEATURES             Location/Qualifiers
     source          1..6584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        866..884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             920..1627
                     /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(1065..1083)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1333..1352
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2178..2195)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2184..2408
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2454..2882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2541..2560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2751..2772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2891..2911)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2896..3225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3076..3211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3138..3157
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3292..4086
                     /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(3346..3365)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3956..3975
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4260..4381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4297..4316)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4351..4370
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4430..4446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4430..4446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4443..4465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4454..4470
                     /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(4478..4484,4485..4502,4503..4508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4523..4544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4661..4678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4832..5420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4912..4931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5591..6382,6383..6451))
                     /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     6214..6233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6452..6556)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      841 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      901 agcgtttaaa cttaagctta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
      961 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     1021 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1081 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     1141 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     1201 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     1261 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1321 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1381 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     1441 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1501 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     1561 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1621 gtacaaggaa ttcatggact cgggccggga cttcctgacc ctgcacggcc tacaggatga
     1681 tgaggatcta caggcgctgc tgaagggcag ccagctcctg aaggtgaagt ccagctcatg
     1741 gaggagagag cgcttctaca agttgcagga ggactgcaag accatctggc aggagtcccg
     1801 caaggtcatg cggaccccgg agtcccagct gttctccatc gaggacattc aggaggtgcg
     1861 aatggggcac cgcacggagg gtctggagaa gttcgcccgt gatgtgcccg aggaccgctg
     1921 cttctccatt gtcttcaagg accagcgcaa tacactagac ctcatcgccc catcgccagc
     1981 tgatgcccag cactgggtgc tggggctgca caagatcatc caccactcag gctccatgga
     2041 ccagcgtcag aagctacagc actggattca ctcctgcttg cgaaaagctg acaaaaacaa
     2101 ggacaacaag atgagcttca aggagctgca gaacttcctg aagtagtcta gagggcccgt
     2161 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2221 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2281 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2341 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2401 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2461 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2521 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2581 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2641 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2701 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2761 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2821 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2881 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3241 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3301 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3361 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3421 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3481 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3541 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3601 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3661 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3721 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3781 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3841 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3901 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3961 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4021 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4081 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4141 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4201 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4381 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5341 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5401 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5461 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5521 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5581 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5641 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5701 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5761 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5821 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5881 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5941 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6001 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6061 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6121 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6181 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6241 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6301 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6361 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6421 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6481 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6541 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgac
//
