LOCUS       sequence-173171-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, N1 Cloning Vector, Excitation:399, 
            Emission:456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5955)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 5955)
  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: mTagBFP2-TES-N-14
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1300..1309
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1306..2016
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    2141..2262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2178..2197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2232..2251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2269..2724)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2406..2427)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2618..2637
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2751..2855
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2857..3214
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2857..3048
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2880..2900)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3065..3200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3249..4043
                     /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             3249..4043
                     /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(3303..3322)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3913..3932
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4231..4250)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4275..4322
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4651..5239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5140..5159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5414..5717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5718..5921
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5872..5892
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5950..5955,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggacctgg aaaacaaagt gaagaagatg ggcttaggtc acgagcaagg atttggagcc
       61 ccttgtttaa aatgcaaaga aaaatgtgaa ggattcgaac tgcacttctg gagaaaaata
      121 tgtcgtaact gcaagtgtgg ccaagaagag catgatgtcc tcttgagcaa tgaagaggat
      181 cgaaaagtgg gaaaactttt tgaagacacc aagtatacca ctctgattgc aaaactaaag
      241 tcagatggaa ttcccatgta taaacgcaat gttatgatat tgacgaatcc agttgctgcc
      301 aagaagaatg tctccatcaa tacagttacc tatgagtggg ctcctcctgt ccagaatcaa
      361 gcattggcca ggcagtacat gcagatgcta cccaaggaaa agcagccagt agcaggctca
      421 gagggggcac agtaccggaa gaagcagctg gcaaagcagc tccctgcaca tgaccaggac
      481 ccttcaaagt gccatgagtt gtctcccaga gaggtgaagg agatggagca gtttgtgaag
      541 aaatataaga gcgaagctct gggagtagga gatgtcaaac ttccctgtga gatggatgcc
      601 caaggcccca aacaaatgaa cattcctgga ggggatagaa gcaccccagc agcagtgggg
      661 gccatggagg acaaatctgc tgagcacaaa agaactcaat attcctgcta ttgctgcaaa
      721 ctgagtatga aagaaggtga cccagccatc tatgccgaaa gggctggcta tgataaactg
      781 tggcacccag cttgttttgt ctgcagcacc tgccatgaac tcctggttga catgatttat
      841 ttttggaaga atgagaagct atactgtggc agacattact gtgacagcga gaaaccccga
      901 tgtgctggct gtgacgagct gatattcagc aatgagtata cccaggcaga aaaccagaat
      961 tggcacctga aacacttctg ctgctttgac tgtgatagca ttctagctgg ggagatatac
     1021 gtgatggtca atgacaagcc cgtgtgcaag ccctgctatg tgaagaatca cgctgtggtg
     1081 tgtcaaggat gccacaatgc catcgaccca gaagtgcagc gggtgaccta taacaatttc
     1141 agctggcatg catccacaga gtgctttctg tgctcttgct gcagcaaatg cctcattggg
     1201 cagaagttca tgccagtaga agggatggtt ttctgttcag tggaatgtaa gaagaggatg
     1261 tcttccggcg gtagcggcgg tagcggggat ccaccggtcg ccaccatggt gtctaagggc
     1321 gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac
     1381 catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
     1441 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc
     1501 ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag
     1561 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
     1621 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc
     1681 agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag
     1741 gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc
     1801 ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca agaccacata tagatccaag
     1861 aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga
     1921 atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac
     1981 tgcgacctcc ctagcaaact ggggcacaag cttaattaag cggccgcgac tctagatcat
     2041 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2101 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2221 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2281 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2341 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2401 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2461 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2521 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2581 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2641 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2701 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2761 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2821 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2881 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2941 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3001 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3061 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3121 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3181 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3241 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3301 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3361 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3421 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3481 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3541 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3601 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3661 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3721 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3781 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3841 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3901 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3961 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4021 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4081 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4141 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4201 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4261 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4321 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4381 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4441 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4501 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4561 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4621 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4681 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4741 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4801 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4861 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4921 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4981 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5041 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5101 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5161 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5221 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5281 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5341 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     5401 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5461 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5521 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5581 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5641 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5701 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5761 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5821 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5881 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     5941 tccgctagcg ccacc
//
