LOCUS       sequence-242412-        5611 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 5611)
  TITLE     2PH-PLCdelta-GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5611)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 2PH-PLCdelta-GFP
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             558..580
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      1491..1500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1497..1499,1500..1502,1503..2216)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1497..1499,1500..1502,1503..2213)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1542..1563)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1803..1822)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2150..2171
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2339..2460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2376..2395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2430..2449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2467..2922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2604..2625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2816..2835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2949..3053
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3055..3412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3055..3246
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3078..3098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3263..3398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3447..4241
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3447..4241
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3501..3520)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4111..4130
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4429..4448)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4473..4520
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4849..5437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5338..5357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctaatttt gtttaacttt aagaaggaga tatacatatg gactcgggcc
      601 gggacttcct gaccctgcac ggcctacagg atgatgagga tctacaggcg ctgctgaagg
      661 gcagccagct cctgaaggtg aagtccagct catggaggag agagcgcttc tacaagttgc
      721 aggaggactg caagaccatc tggcaggagt cccgcaaggt catgcggacc ccggagtccc
      781 agctgttctc catcgaggac attcaggagg tgcgaatggg gcaccgcacg gagggtctgg
      841 agaagttcgc ccgtgatgtg cccgaggacc gctgcttctc cattgtcttc aaggaccagc
      901 gcaatacact agacctcatc gccccatcgc cagctgatgc ccagcactgg gtgctggggc
      961 tgcacaagat catccaccac tcaggctcca tggaccagcg tcagaagcta cagcactgga
     1021 ttcactcctg cttgcgaaaa ggtggcaaaa acaaggacaa caagataatc catatggact
     1081 cgggccggga cttcctgacc ctgcacggcc tacaggatga tgaggatcta caggcgctgc
     1141 tgaagggcag ccagctcctg aaggtgaagt ccagctcatg gaggagagag cgcttctaca
     1201 agttgcagga ggactgcaag accatctggc aggagtcccg caaggtcatg cggaccccgg
     1261 agtcccagct gttctccatc gaggacattc aggaggtgcg aatggggcac cgcacggagg
     1321 gtctggagaa gttcgcccgt gatgtgcccg aggaccgctg cttctccatt gtcttcaagg
     1381 accagcgcaa tacactagac ctcatcgccc catcgccagc tgatgcccag cactgggtgc
     1441 tggggctgca caagatcatc caccactcag gctccatgga tccaccggtc gccaccatgg
     1501 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1561 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1621 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1681 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1741 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1801 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1861 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1921 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1981 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2041 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2101 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2161 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     2221 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2281 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2341 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2401 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2461 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2941 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3001 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3061 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3421 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3481 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3541 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3601 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3661 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3721 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3781 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3841 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3901 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3961 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4021 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4081 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4141 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4201 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4261 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4321 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4381 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4441 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4501 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4561 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4621 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4681 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4981 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5521 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     5581 cattagttca tagcccatat atggagttcc g
//
