LOCUS       sequence-442804-        6399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse EGFP-Paxillin .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6399)
  AUTHORS   Chinten James Lim
  TITLE     Chinten James Lim
  JOURNAL   Chinten James Lim
REFERENCE   2  (bases 1 to 6399)
  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: "Chinten 
            James Lim"
COMMENT     Sequence Label: EGFP-m-Paxillin
FEATURES             Location/Qualifiers
     source          1..6399
                     /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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3127..3248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3164..3183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3218..3237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3255..3710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3392..3413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3604..3623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3737..3841
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3843..4200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3843..4034
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3866..3886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4051..4186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4235..5029
                     /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             4235..5029
                     /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(4289..4308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4899..4918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5217..5236)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5261..5308
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5637..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attcagacga cctcgatgcc
     1321 ctgctggcag acttggaatc taccacctcc cacatctcca aacggccagt gttcttgtca
     1381 gaggagcccc cctactccta cccaactgga aaccacacat accaggagat tgcggtgcca
     1441 cctccggtcc caccaccacc gtccagcgag gccctcaatg gcacggtcct ggacccttta
     1501 gatcagtggc agcctagtgg ttccagatac gctcaccagc agcctccgtc cccactgccc
     1561 gtgtacagct ccagtgctaa aaattccagt gcctccaaca cccaggacgg cgtgggctcc
     1621 ctttgttccc gagcaggcga ggaagagcac gtctacagct tccccaacaa gcagaagtcg
     1681 gcagagccct cacctaccgt catgagctcc tccctgggca gcaacctctc tgaacttgac
     1741 cggctgttac tggagctgaa cgcagtgcag cacagcccgc ctggcttccc tgcagatgaa
     1801 gccgaatcga gccccccctt gcctggagcc ctgagccccc tttatggcat cccagaaaat
     1861 aacactccct tggggggcaa agcggggccc ctggtgaaag agaagccaaa gcgaaatgga
     1921 ggccgggggc tggaggatgt gcggcccagc gtggagagcc tcttggatga actggagagt
     1981 tccgtgccca gccctgtccc ggccatcact gtgaaccagg gagagatgag cagtccgcag
     2041 cgagtcacct ccagccagca gcagacccgg atctcggcct cctctgccac cagggagctg
     2101 gacgagctca tggcgtcact gtcagatttc aagttcatgg cccaggggaa aactgggagc
     2161 agctcacccc ctgggggact ctccaagccc gggagccagc tggacagcat gttggggagt
     2221 ctgcagtccg acctgaacaa gctaggggtc gccaccgttg ccaaaggggt ctgtggagcc
     2281 tgcaagaagc ccatcgccgg gcaggtcgtg accgccatgg ggaagacatg gcaccccgag
     2341 cactttgtgt gcacccactg ccaggaggag attggatccc gaaacttttt tgagcgggac
     2401 ggacagccct actgtgagaa ggactaccac agcctcttct ccccacgctg ctactactgc
     2461 aacggaccca tcctggataa agtggtgaca gcgctggacc gcacatggca cccggaacac
     2521 ttcttctgtg cccagtgtgg agccttcttt ggtccagaag gcttccacga gaaggacggc
     2581 aaagcgtact gtcgtaaaga ttacttcgac atgtttgctc ccaagtgcgg cggctgcgcc
     2641 cgtgccatcc tggagaacta catttcagcc ctcaacaccc tctggcaccc cgaatgcttt
     2701 gtgtgcaggg agtgcttcac acccttcgtc aacggcagct tctttgagca cgacgggcag
     2761 ccgtactgtg aggtccacta ccacgagcgg cgcggctcgc tgtgctccgg ctgccagaag
     2821 cccatcacgg gccgctgcat caccgccatg gccaagaagt tccaccccga gcatttcgtg
     2881 tgcgcctttt gcctgaagca gctcaacaag ggcaccttca aggagcagaa cgacaaaccc
     2941 tactgccaga gctgcttcgt gaagctcttc tgctaggtcg acggtaccgc gggcccggga
     3001 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3061 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3121 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3241 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3301 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3361 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3421 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3481 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3541 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3601 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3661 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3721 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3781 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3841 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4021 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4201 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4261 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4321 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4381 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4441 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4501 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4561 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4621 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4681 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4741 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4801 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4861 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4921 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4981 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5041 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5101 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5161 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5221 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5281 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5341 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5401 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5461 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5521 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5581 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6301 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     6361 tacggggtca ttagttcata gcccatatat ggagttccg
//
