pcDNA3-GRITOL

LOCUS       pcDNA3-GRITOL               6602 bp DNA     circular SYN 17-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6602)
  AUTHORS   tao
  TITLE     Direct Submission
  JOURNAL   Exported Jul 6, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..906
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    907..1980
                     /label=TrpR sensor
     CDS             907..1098
                     /codon_start=1
                     /label=TrpR-N
                     /translation="MAQQSPYSAAMAEQRHQEWLRFVDLLKNAYQNDLHLPLLNLMLTP
                     DEREALGTGVRIVEELLRG"
     CDS             1099..1836
                     /codon_start=1
                     /label=cpsfYFP
                     /translation="FNSDNVYIMADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIG
                     DGPVLLPDNHYLSFQSMLSKDPNEKRDHMVLLEFVTAAGITLGMDELYNVDGGSGGTGS
                     KGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKLICTTGKLPVPWTTLV
                     TTLGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGTYKTRAEVKFEGDTLV
                     NRIELKGIGFKEDGNILGHKLEYN"
     misc_feature    1282..1284
                     /label=GTC->ATG
     CDS             1843..1971
                     /codon_start=1
                     /label=TrpR-C
                     /translation="MSQEELKNELGAGIATITRGSNSLKAAPVELRQWLEEVLLKSD"
     misc_feature    1981..2015
                     /label=Standard pcDNA 3-MCS site
     polyA_signal    2033..2257
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2303..2731
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2745..3074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2925..3060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3123..4148
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4278..4399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4448..4464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4472..4488
                     /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(4496..4526)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4541..4562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4850..5435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5606..6466)
                     /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"
     promoter        complement(6467..6571)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg cccaacaatc accctattca gcagcgatgg cagaacagcg tcaccaggag
      961 tggttacgtt ttgtcgacct gcttaagaat gcctaccaaa acgatctcca tttaccgttg
     1021 ttaaacctga tgctgacgcc agatgagcgc gaagcgttgg ggactggcgt gcgtattgtg
     1081 gaagagctgt tgcgcggctt caacagcgac aacgtctata tcatggccga caagcagaag
     1141 aacggcatca aggccaactt caagatccgc cacaacgtcg aggacggcag cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gcttccagtc catgctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaac
     1381 gtggatggcg gtagcggtgg caccggcagc aagggcgagg agctgttcac cggggtggtg
     1441 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgtggcgag
     1501 ggcgagggcg atgccaccaa cggcaagctg accctgaagc tgatctgcac caccggcaag
     1561 ctgcccgtgc cctggaccac cctcgtgacc accctcggct acggcctgaa gtgcttcgcc
     1621 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1681 gtccaggagc gcaccatctt cttcaaggac gacggcactt acaagacccg cgccgaggtg
     1741 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcgg cttcaaggag
     1801 gacggcaaca tcctggggca caagctggag tacaaccagg ccatgagcca ggaggagtta
     1861 aaaaatgaac tcggcgcggg catcgcgacg attacgcgtg gatctaacag cctgaaagcc
     1921 gcgcctgtcg agctgcgcca gtggctggaa gaggtgttgc tgaaaagcga tctagcataa
     1981 aagcttaagc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2041 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2101 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2161 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2221 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2281 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2341 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2401 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2461 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2521 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2581 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2641 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2701 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2821 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2881 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2941 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3001 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3061 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     3121 tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     3181 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     3241 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     3301 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     3361 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     3421 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     3481 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     3541 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     3601 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     3661 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     3721 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     3781 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     3841 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     3901 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     3961 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     4021 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     4081 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     4141 aggaatagca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4201 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4261 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4381 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5401 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5521 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5581 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5641 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5701 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5761 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5821 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5881 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5941 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6001 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6061 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6121 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6181 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6241 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6301 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6361 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6421 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6481 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6541 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6601 tc
//