LOCUS       sequence-216427-        7893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 492, Emission: 505.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7893)
  TITLE     Davidson Green fluorescent proteins
REFERENCE   2  (bases 1 to 7893)
  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"
COMMENT     Sequence Label: mAzami Green-Vinculin-23
FEATURES             Location/Qualifiers
     source          1..7893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             671..1348
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             671..1348
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     polyA_signal    4740..4861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4777..4796)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4831..4850
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4868..5323)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5005..5026)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5217..5236
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5350..5454
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5456..5813
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5456..5647
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5479..5499)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5664..5799
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5847..6641
                     /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             5847..6641
                     /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(5901..5920)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6511..6530
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6829..6848)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6873..6920
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7249..7837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7738..7757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagcg tgatcaagcc cgagatgaag atcaagctgt gcatgagggg
      721 caccgtgaac ggccacaact tcgtgatcga gggcgagggc aagggcaacc cctacgaggg
      781 cacccagatc ctggacctga acgtgaccga gggcgccccc ctgcccttcg cctacgacat
      841 cctgaccacc gtgttccagt acggcaacag ggccttcacc aagtaccccg ccgacatcca
      901 ggactacttc aagcagacct tccccgaggg ctaccactgg gagaggagca tgacctacga
      961 ggaccagggc atctgcaccg ccaccagcaa catcagcatg aggggcgact gcttcttcta
     1021 cgacatcagg ttcgacggca ccaacttccc ccccaacggc cccgtgatgc agaagaagac
     1081 cctgaagtgg gagcccagca ccgagaagat gtacgtggag gacggcgtgc tgaagggcga
     1141 cgtgaacatg aggctgctgc tggagggcgg cggccactac aggtgcgact tcaagaccac
     1201 ctacaaggcc aagaaggagg tgaggctgcc cgacgcccac aagatcgacc acaggatcga
     1261 gatcctgaag cacgacaagg actacaacaa ggtgaagctg tacgagaacg ccgtggccag
     1321 gtactccatg ctgcccagcc aggccaagtc cggactcaga tctcgagctc aagcttcgaa
     1381 ttcgggaaac tcgagaacaa agcttgaaaa cggccgcatg ccagtgtttc atacgcgcac
     1441 gatcgagagc atcctggagc cggtggcaca gcagatctcc cacctggtga taatgcacga
     1501 ggagggcgag gtggacggca aagccattcc tgacctcacc gcgcccgtgg ccgccgtgca
     1561 ggcggccgtc agcaacctcg tccgggttgg aaaagagact gttcaaacca ctgaggatca
     1621 gattttgaag agagatatgc caccagcatt tattaaggtt gagaatgctt gcaccaagct
     1681 tgtccaggca gctcagatgc ttcagtcaga cccttactca gtgcctgctc gagattatct
     1741 aattgatggg tcaaggggca tcctctctgg aacatcagac ctgctcctta ccttcgatga
     1801 ggctgaggtc cgtaaaatta ttagagtttg caaaggaatt ttggaatatc ttacagtggc
     1861 agaggtggtg gagactatgg aagatttggt cacttacaca aagaatcttg ggccaggaat
     1921 gactaagatg gccaagatga ttgacgagag acagcaggag ctcactcacc aggagcaccg
     1981 agtgatgttg gtgaactcga tgaacaccgt gaaagagttg ctgccagttc tcatttcagc
     2041 tatgaagatt tttgtaacaa ctaaaaactc aaaaaaccaa ggcatagagg aagctttaaa
     2101 aaatcgcaat tttactctag aaaaaatgag tgctgaaatt aatgagataa ttcgtgtgtt
     2161 acaactcacc tcttgggatg aagatgcctg ggccagcaag gacactgaag ccatgaagag
     2221 agcattggcc tccatagact ccaaactgaa ccaggccaaa ggttggctcc gtgaccctag
     2281 tgcctcccca ggggatgctg gtgagcaggc catcagacag atcttagatg aagctggaaa
     2341 agttggtgaa ctctgtgcag gcaaagaacg cagggagatt ctgggaactt gcaaaatgct
     2401 agggcagatg actgatcaag tggctgacct ccgtgccaga ggacaaggat cctcaccggt
     2461 ggccatgcag aaagctcagc aggtatctca gggtctggat gtgctcacag caaaagtgga
     2521 aaatgcagct cgcaagctgg aagccatgac caactcaaag cagagcattg caaagaagat
     2581 cgatgctgct cagaactggc ttgcagatcc aaatggtgga ccggaaggag aagagcagat
     2641 tcgaggtgct ttggctgaag ctcggaaaat agcagaatta tgtgatgatc ctaaagaaag
     2701 agatgacatt ctacgttccc ttggggaaat atctgctctg acttctaaat tagcagatct
     2761 acgaagacag gggaaaggag attctccaga ggctcgagcc ttggccaaac aggtggccac
     2821 ggccctgcag aacctgcaga ccaaaaccaa ccgggctgtg gccaacagca gaccggccaa
     2881 agcagctgta caccttgagg gcaagattga gcaagcacag cggtggattg ataatcccac
     2941 agtggatgac cgtggagtcg gtcaggctgc catccggggg cttgtggccg aagggcatcg
     3001 tctggctaat gttatgatgg ggccttatcg gcaagatctt ctcgccaagt gtgaccgagt
     3061 ggaccagctg acagcccagc tggctgacct ggctgccaga ggggaagggg agagtcctca
     3121 ggcacgagca cttgcatctc agctccaaga ctccttaaag gatctaaaag ctcggatgca
     3181 ggaggccatg actcaggaag tgtcagatgt tttcagcgat accacaactc ccatcaagct
     3241 gttggcagtg gcagccacgg cgcctcctga tgcgcctaac agggaagagg tatttgatga
     3301 gagggcagct aactttgaaa accattcagg aaagcttggt gctacggccg agaaggcggc
     3361 tgcggttggt actgctaata aatcaacagt ggaaggcatt caggcctcag tgaagacggc
     3421 ccgagaactc acaccccagg tggtctcggc tgctcgtatc ttacttagga accctggaaa
     3481 tcaagctgct tatgaacatt ttgagaccat gaagaaccag tggatcgata atgttgaaaa
     3541 aatgacaggg ctggtggacg aagccattga taccaaatct ctgttggatg cttcagaaga
     3601 agcaattaaa aaagacctgg acaagtgcaa ggtagctatg gccaacattc agcctcagat
     3661 gctggttgct ggggcaacca gtattgctcg tcgggccaac cggatcctgc tggtggctaa
     3721 gagggaggtg gaaaattccg aggatcccaa gttccgtgag gctgtgaaag ctgcctctga
     3781 tgaattgagc aaaaccatct ccccgatggt gatggatgca aaagctgtgg ctggaaacat
     3841 ttccgaccct ggactgcaaa agagcttcct ggactcagga tatcggatcc tgggagctgt
     3901 ggccaaggtc agagaagcct tccaacctca ggagcctgac ttcccgccgc ctccaccaga
     3961 ccttgaacaa ctccgactaa cagatgagct tgctcctccc aaaccacctc tgcctgaagg
     4021 tgaggtccct ccacctaggc ctccaccacc agaggaaaag gatgaagagt tccctgagca
     4081 gaaggccggg gaggtgatta accagccaat gatgatggct gccagacagc tccatgatga
     4141 agctcgcaaa tggtccagca agggcaatga catcattgca gcagccaagc gcatggctct
     4201 gctgatggct gagatgtctc ggctggtaag agggggcagt ggtaccaagc gggcactcat
     4261 tcagtgtgcc aaggacatcg ccaaggcctc agatgaggtg actcggttgg ccaaggaggt
     4321 tgccaagcag tgcacagata aacggattag aaccaacctc ttacaggtat gtgagcgaat
     4381 cccaaccata agcacccagc tcaaaatcct gtccacagtg aaggccacca tgctgggccg
     4441 gaccaacatc agtgatgagg agtctgagca ggccacagag atgctggttc acaatgccca
     4501 gaacctcatg cagtctgtga aggagactgt gcgggaagct gaagctgctt caatcaaaat
     4561 tcgaacagat gctggattta cactgcgctg ggttagaaag actccctggt accagtaggc
     4621 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     4681 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4741 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4801 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4861 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4921 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4981 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5041 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5101 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5161 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5221 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5281 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     5341 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5401 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     5461 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5581 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5641 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5701 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5761 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcatc
     5821 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5881 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5941 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     6001 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     6061 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     6121 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     6181 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     6241 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6301 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6361 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6421 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6481 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6541 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6601 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6661 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6721 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6781 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6841 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6901 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6961 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     7021 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     7081 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7141 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7201 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7861 acggttcctg gccttttgct ggccttttgc tca
//
