pUBC-3N75-EGFP

LOCUS       pUBC-3N75-EGFP         7275 bp DNA     circular SYN 11-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7275)
  AUTHORS   Ruizhao Wang
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      55..60
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             61..2205
                     /codon_start=1
                     /label=3N75(D460L)
                     /translation="MNVIAILNHMGVYFKEEPIRELHRALERLNFQIVYPNDRDDLLKL
                     IENNARLCGVIFDWDKYNLELCEEISKMNENLPLYAFANTYSTLDVSLNDLRLQISFFE
                     YALGAAEDIANKIKQTTDEYINTILPPLTKALFKYVREGKYTFCTPGHMGGTAFQKSPV
                     GSLFYDFFGPNTMKSDISISVSELGSLLDHSGPHKEAEQYIARVFNADRSYMVTNGTST
                     ANKIVGMYSAPAGSTILIDRNCHKSLTHLMMMSDVTPIYFRPTRNAYGILGGIPQSEFQ
                     HATIAKRVKETPNATWPVHAVITNSTYDGLLYNTDFIKKTLDVKSIHFDSAWVPYTNFS
                     PIYEGKCGMSGGRVEGKVIYETQSTHKLLAAFSQASMIHVKGDVNEETFNEAYMMHTTT
                     SPHYGIVASTETAAAMMKGNAGKRLINGSIERAIKFRKEIKRLRTESDGWFFDVWQPDH
                     ILTTECWPLRSDSTWHGFKNIDNEHMYLDPIKVTLLTPGMEKDGTMSDFGIPASIVAKY
                     LDEHGIVVEKTGPYNLLFLFSIGIDKTKALSLLRALTDFKRAFDLNLRVKNMLPSLYRE
                     DPEFYENMRIQELAQNIHKLIVHHNLPDLMYRAFEVLPTMVMTPYAAFQKELHGMTEEV
                     YLDEMVGRINANMILPYPPGVPLVMPGEMITEESRPVLEFLQMLCEIGAHYPGFETDIH
                     GAYRQADGRYTVKVLKEESKK"
     CDS             2206..2235
                     /codon_start=1
                     /label=Linker10
                     /translation="GGGGSGGGGS"
     CDS             2237..2952
                     /codon_start=1
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     polyA_signal    3104..3159
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3520..3536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3544..3560
                     /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(3568..3598)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     3573..3594
                     /label=lac-pro-F
     protein_bind    3613..3634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3693..3888
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3718..3888
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3739..3874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3894..4028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4267..4855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4271..4290)
                     /label=pUBC-primer-1
     CDS             complement(5026..5886)
                     /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     complement(5057..5078)
                     /label=Amp-F
     primer_bind     5800..5820
                     /label=Amp-R-seq
     promoter        complement(5887..5990)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6063..6462
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(6190..6212)
                     /label=hUBC-pro-R
     primer_bind     6658..6681
                     /label=pUBC-seq-2
     promoter        6749..7275
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7112..7131
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgccacc
       61 atgaacgtga tcgccatcct gaaccacatg ggcgtgtact tcaaggagga gcccatcagg
      121 gagctgcaca gggccctgga gaggctgaac ttccagatcg tgtaccccaa cgacagggac
      181 gacctgctga agctgatcga gaacaacgcc aggctgtgcg gcgtgatctt cgactgggac
      241 aagtacaacc tggagctgtg cgaggagatc agcaagatga acgagaacct gcccctgtac
      301 gccttcgcca acacctacag caccctggac gtgagcctga acgacctgag gctgcagatc
      361 agcttcttcg agtacgccct gggcgccgcc gaggacatcg ccaacaagat caagcagacc
      421 accgacgagt acatcaacac catcctgccc cccctgacca aggccctgtt caagtacgtg
      481 agggagggca agtacacctt ctgcaccccc ggccacatgg gcggcaccgc cttccagaag
      541 agccccgtgg gcagcctgtt ctacgacttc ttcggcccca acaccatgaa gagcgacatc
      601 agcatcagcg tgagcgagct gggcagcctg ctggaccaca gcggccccca caaggaggcc
      661 gagcagtaca tcgccagggt gttcaacgcc gacaggagct acatggtgac caacggcacc
      721 agcaccgcca acaagatcgt gggcatgtac agcgcccccg ccggcagcac catcctgatc
      781 gacaggaact gccacaagag cctgacccac ctgatgatga tgagcgacgt gacccccatc
      841 tacttcaggc ccaccaggaa cgcctacggc atcctgggcg gcatccccca gagcgagttc
      901 cagcacgcca ccatcgccaa gagggtgaag gagaccccca acgccacctg gcccgtgcac
      961 gccgtgatca ccaacagcac ctacgacggc ctgctgtaca acaccgactt catcaagaag
     1021 accctggacg tgaagagcat ccacttcgac agcgcctggg tgccctacac caacttcagc
     1081 cccatctacg agggcaagtg cggcatgagc ggcggcaggg tggagggcaa ggtgatctac
     1141 gagacccaga gcacccacaa gctgctggcc gccttcagcc aggccagcat gatccacgtg
     1201 aagggcgacg tgaacgagga gaccttcaac gaggcctaca tgatgcacac caccaccagc
     1261 ccccactacg gcatcgtggc cagcaccgag accgccgccg ccatgatgaa gggcaacgcc
     1321 ggcaagaggc tgatcaacgg cagcatcgag agggccatca agttcaggaa ggagatcaag
     1381 aggctgagga ccgagagcga cggctggttc ttcgacgtgt ggcagcccga ccacatcctg
     1441 accaccgagt gctggcccct gaggagcgac agcacctggc acggcttcaa gaacatcgac
     1501 aacgagcaca tgtacctgga ccccatcaag gtgaccctgc tgacccccgg catggagaag
     1561 gacggcacca tgagcgactt cggcatcccc gccagcatcg tggccaagta cctggacgag
     1621 cacggcatcg tggtggagaa gaccggcccc tacaacctgc tgttcctgtt cagcatcggc
     1681 atcgacaaga ccaaggccct gagcctgctg agggccctga ccgacttcaa gagggccttc
     1741 gacctgaacc tgagggtgaa gaacatgctg cccagcctgt acagggagga ccccgagttc
     1801 tacgagaaca tgaggatcca ggagctggcc cagaacatcc acaagctgat cgtgcaccac
     1861 aacctgcccg acctgatgta cagggccttc gaggtgctgc ccaccatggt gatgaccccc
     1921 tacgccgcct tccagaagga gctgcacggc atgaccgagg aggtgtacct ggacgagatg
     1981 gtgggcagga tcaacgccaa catgatcctg ccctaccccc ccggcgtgcc cctggtgatg
     2041 cccggcgaga tgatcaccga ggagagcagg cccgtgctgg agttcctgca gatgctgtgc
     2101 gagatcggcg cccactaccc cggcttcgag accgacatcc acggcgccta caggcaggcc
     2161 gacggcaggt acaccgtgaa ggtgctgaag gaggagagca agaagggcgg cggcggcagc
     2221 ggcggcggcg gcagcgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2281 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2341 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2401 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2461 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2521 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2581 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2641 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2701 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2761 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2821 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2881 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2941 ctgtacaagt aaagcggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     3001 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     3061 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     3121 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     3181 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     3241 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     3301 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     3361 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     3421 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     3481 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3661 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     3721 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3781 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3841 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     3901 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3961 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4021 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4081 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4201 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4261 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4321 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4381 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4441 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4501 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4561 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4621 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4681 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4741 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4801 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4861 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4921 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4981 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5041 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5101 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5161 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5221 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5281 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5341 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5401 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5461 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5521 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5581 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5641 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5701 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5761 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5821 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5881 actcatactc ttcctttttc atattattga agcatttatc agggttattg tctcatgagc
     5941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6001 cgaaaagtgc cacctgggtc gacattgatt attgactagt aacccgtgtc ggctccagat
     6061 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc
     6121 tgccacgtca gacgaagggc gcagcgagcg tcctgatcct tccgcccgga cgctcaggac
     6181 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     6241 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     6301 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     6361 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     6421 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     6481 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     6541 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggga
     6601 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     6661 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgaggc
     6721 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc atctggggac
     6781 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     6841 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     6901 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     6961 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     7021 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     7081 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     7141 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     7201 tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg gccgtttttg
     7261 gcttttttgt tagac
//