pCDH_GFP_LplA

LOCUS       pCDH_GFP_LplA               8184 bp DNA     circular SYN 03-JUL-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8184)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Aug 24, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    45..170
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    663..896
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1392..1508
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1531..1834
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1835..2038
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2083..2092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2089..2805
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2806..2829
                     /codon_start=1
                     /label=linker
                     /translation="GGGSGGGS"
     CDS             2830..3843
                     /codon_start=1
                     /product="E. coli LplA"
                     /label=LplA
                     /translation="STLRLLISDSYDPWFNLAVEECIFRQMPATQRVLFLWRNADTVVI
                     GRAQNPWKECNTRRMEEDNVRLARRSSGGGAVFHDLGNTCFTFMAGKPEYDKTISTSIV
                     LNALNALGVSAEASGRNDLVVKTVEGDRKVSGSAYRETKDRGFHHGTLLLNADLSRLAN
                     YLNPDKKKLAAKGITSVRSRVTNLTELLPGITHEQVCEAITEAFFAHYGERVEAEIISP
                     NKTPDLPNFAETFARQSSWEWNFGQAPAFSHLLDERFTWGGVELHFDVEKGHITRAQVF
                     TDSLNPAPLEALAGRLQGCLYRADMLQQECEALLVDFPEQEKELRELSAWMAGAVR"
     misc_feature    3850..4437
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4321..4332)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4510..4743
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4815..4936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4976..5111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5144..5160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5168..5184
                     /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(5192..5222)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5237..5258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5546..6134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6305..7165)
                     /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(7166..7270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7744..7760
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7775..8001
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8002..8182
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
       61 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      121 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      181 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
      241 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
      301 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
      361 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
      421 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
      481 acaaaagtaa gaccaccgca cagcaagcgg ccactgatct tcagacctgg aggaggagat
      541 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
      601 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
      661 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
      721 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
      781 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
      841 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
      901 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
      961 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1021 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1081 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1141 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1201 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     1261 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     1321 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     1381 tatcggttaa cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     1441 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc
     1501 aaaattttat cgatactagt tggagttccg cgttacataa cttacggtaa atggcccgcc
     1561 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1621 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1681 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1741 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1801 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1861 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1921 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1981 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     2041 tttagtgaac cgtcagattc tagaccgcta gcgctaccgg tcgccaccat ggtgagcaag
     2101 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2161 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2221 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2281 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2341 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2401 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2461 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2521 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2581 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2641 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2701 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2761 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg cggttccggc
     2821 ggcggcagct ccacattacg cctgctcatc tctgactctt acgacccgtg gtttaacctg
     2881 gcggtggaag agtgtatttt tcgccaaatg cccgccacgc agcgcgttct gtttctctgg
     2941 cgcaatgccg acacggtagt aattggtcgc gcgcagaacc cgtggaaaga gtgtaatacc
     3001 cggcggatgg aagaagataa cgtccgcctg gcgcggcgca gtagcggtgg cggcgcggtg
     3061 ttccacgatc tcggcaatac ctgctttacc tttatggctg gcaagccgga gtacgataaa
     3121 actatctcca cgtcgattgt gctcaatgcg ctgaacgcgc tcggcgtcag cgccgaagcg
     3181 tccggacgta acgatctggt ggtgaaaacc gtcgaaggcg accgcaaagt ctcaggctcg
     3241 gcctatcgcg aaaccaaaga tcgcggcttc caccacggca ccttgctact caatgccgac
     3301 ctcagccgcc tggcaaacta tctcaatccg gataaaaaga aactggcggc gaaaggcatt
     3361 acgtcggtac gttcccgcgt gaccaacctc accgagctgt tgccggggat cacccatgag
     3421 caggtttgcg aggccataac cgaggccttt ttcgcccatt atggcgagcg cgtggaagcg
     3481 gaaatcatct ccccgaacaa aacgccagac ttgccaaact tcgccgaaac ctttgcccgc
     3541 cagagtagct gggaatggaa cttcggtcag gctccggcat tctcgcatct gctggatgaa
     3601 cgctttacct ggggcggcgt ggaactgcat ttcgacgttg aaaaaggcca tatcacccgc
     3661 gcccaggtgt ttaccgacag cctcaacccc gcgccgctgg aagccctcgc cggacgactg
     3721 caaggctgcc tgtaccgcgc agatatgctg caacaggagt gcgaagcgct gttggttgac
     3781 ttcccggaac aggaaaaaga gctacgggag ttatcggcat ggatggcggg ggctgtaagg
     3841 tagggatcca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3901 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3961 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4021 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4081 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4141 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4201 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     4261 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4321 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4381 ccgcgtctcc gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgccgg
     4441 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     4501 aggggggact ggaagggcta attcactccc aacgaagata agatctgctt tttgcttgta
     4561 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     4621 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     4681 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     4741 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     4801 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4861 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4921 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     4981 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5041 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5101 ttttggaggc ctagactttt gcagagacca aattcgtaat catgtcatag ctgtttcctg
     5161 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5221 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5281 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5341 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5521 ttaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5581 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5641 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5701 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5761 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5821 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5881 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5941 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6001 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6061 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6121 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6181 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6241 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7261 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7321 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7381 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7441 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7501 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7561 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     7621 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     7681 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     7741 gttgtaaaac gacggccagt gccaagctga cgcgtgtagt cttatgcaat actcttgtag
     7801 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     7861 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     7921 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     7981 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     8041 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     8101 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     8161 tcagtgtgga aaatctctag cagt
//