LOCUS       sequence-323960-        6048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Talin Head Domain, H, Excitation:487, Emission:509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 6048)
  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: mEmerald Talin-H-18
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..317
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        318..521
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     472..492
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      560..569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(566..568,569..571,572..1282)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(611..632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(872..891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1219..1240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1331..1340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2789..2910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2826..2845)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2880..2899
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2917..3372)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3054..3075)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3266..3285
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3399..3503
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3505..3862
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3505..3696
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3528..3548)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3713..3848
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3897..4691
                     /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             3897..4691
                     /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(3951..3970)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4561..4580
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4879..4898)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4923..4970
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5299..5887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5788..5807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
       61 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      301 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      361 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      421 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      481 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      541 tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      601 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      661 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      721 caagctgccc gtgccctggc ccaccctcgt gaccaccttg acctacggcg tgcagtgctt
      781 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      841 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      901 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      961 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta
     1021 tatcaccgcc gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat
     1081 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1141 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     1201 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1261 cggcatggac gagctgtaca agtccggact cagatctcga gctcaagcta gcttcgaatt
     1321 cgcagaagct gccaccatgg ttgcgctttc gctgaagatt agcattggga atgtggtgaa
     1381 gacgatgcaa tttgagccat ctaccatggt gtacgacgcc tgccgcatga ttcgtgagcg
     1441 gatcccagag gccctggctg gccctcccaa cgactttggg ctctttctgt cagatgatga
     1501 ccccaaaaaa ggcatctggc tagaggctgg gaaagctttg gactactaca tgctccgaaa
     1561 tggggacacc atggagtaca gaaaaaagca gagacccctg aagatccgga tgttagacgg
     1621 aacggtgaag actatcatgg tggatgactc caagaccgtc acagacatgc tcatgaccat
     1681 ttgtgcccga attggtatca ccaaccatga tgagtattca ctggttcgag agctgatgga
     1741 agaaaagaaa gatgagggga cagggaccct gagaaaagac aagaccctac tgcgagatga
     1801 aaagaagatg gagaagctaa agcagaagtt gcacacagac gatgagttga actggctgga
     1861 ccatgggcgg acactgaggg aacagggagt ggaagagcat gagacgttgc tgctgcggag
     1921 aaagttcttc tactcagacc agaatgtgga ttcccgagac cctgtacagc tgaaccttct
     1981 ctatgtgcag gcacgagatg acatcctgaa tggctcccat cctgtctcct ttgacaaggc
     2041 ctgtgaattt gcaggcttcc agtgccagat ccagtttgga cctcacaatg aacagaagca
     2101 caaggctggc ttccttgacc tgaaggactt cctgcccaag gagtatgtga agcagaaggg
     2161 agagcgtaag atctttcagg cacacaagaa ttgtgggcag atgagtgaaa ttgaggccaa
     2221 ggtacgctac gtgaagctgg cccgttccct caagacttac ggtgtctcct tcttcctagt
     2281 caaggaaaag atgaagggga agaataaact ggtgcccagg ctgttgggca tcactaagga
     2341 gtgtgtgatg cgtgtggatg agaagaccaa ggaggtgatt caggaatgga gcctcaccaa
     2401 catcaaacgc tgggctgcct ctcccaagag ctttactctg gactttggag actatcagga
     2461 tggctactac tcagtacaga cgacagaagg cgagcagatc gcacagctca ttgctggcta
     2521 catagatatc atccttaaga agaaaaaaag caaggaccat tttgggctgg agggagatga
     2581 agagtctact atgttggagg actcagtgtc tcccaaaaag tcaacagtcc ttcagtaagt
     2641 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     2701 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2761 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2821 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2881 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     2941 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3001 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3061 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3121 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3181 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3241 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3301 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3361 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3481 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3541 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3781 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3841 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3901 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3961 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4021 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4081 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4141 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4201 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4261 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4321 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4381 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4441 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4501 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4561 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4621 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4681 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4741 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4801 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4861 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4921 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4981 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5041 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5101 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5161 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5221 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5281 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5341 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5401 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5461 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5521 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5581 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5641 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5701 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5761 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5821 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5881 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5941 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6001 ttagttatta atagtaatca attacggggt cattagttca tagcccat
//
