LOCUS       sequence-237123-        7876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 462, Emission: 492.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7876)
  AUTHORS   Ai HW, Olenych SG, Wong P, Davidson MW, Campbell RE
  TITLE     Hue-shifted monomeric variants of Clavularia cyan fluorescent 
            protein: identification of the molecular determinants of color and 
            applications in fluorescence imaging.
  JOURNAL   BMC Biol. 2008 Mar 6;6:13. doi: 10.1186/1741-7007-6-13.
  PUBMED    18325109
REFERENCE   2  (bases 1 to 7876)
  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: "BMC Biol.";
            date: "2008-03-6"; volume: "6:13. doi"; pages: " 
            10.1186/1741-7007-6-13"
COMMENT     Sequence Label: mTFP1-Alpha-V-Integrin-25
FEATURES             Location/Qualifiers
     source          1..7876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        413..616
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3812..3864
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      3874..3883
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3880..4590
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    4712..4833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4749..4768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4803..4822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4840..5295)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4977..4998)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5189..5208
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5322..5426
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5428..5785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5428..5619
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5451..5471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5636..5771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5820..6614
                     /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             5820..6614
                     /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(5874..5893)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6484..6503
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6802..6821)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6846..6893
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7222..7810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7711..7730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagtcgacgg tacctcggcg
      661 atggcttttc cgccgcggcg acggctgcgc ctcggtcccc gcggcctccc gcttcttctc
      721 tcgggactcc tgctacctct gtgccgcgcc ttcaacctag acgtggacag tcctgccgag
      781 tactctggcc ccgagggaag ttacttcggc ttcgccgtgg atttcttcgt gcccagcgcg
      841 tcttcccgga tgtttcttct cgtgggagct cccaaagcaa acaccaccca gcctgggatt
      901 gtggaaggag ggcaggtcct caaatgtgac tggtcttcta cccgccggtg ccagccaatt
      961 gaatttgatg caacaggcaa tagagattat gccaaggatg atccattgga atttaagtcc
     1021 catcagtggt ttggagcatc tgtgaggtcg aaacaggata aaattttggc ctgtgcccca
     1081 ttgtaccatt ggagaactga gatgaaacag gagcgagagc ctgttggaac atgctttctt
     1141 caagatggaa caaagactgt tgagtatgct ccatgtagat cacaagatat tgatgctgat
     1201 ggacagggat tttgtcaagg aggattcagc attgatttta ctaaagctga cagagtactt
     1261 cttggtggtc ctggtagctt ttattggcaa ggtcagctta tttcggatca agtggcagaa
     1321 atcgtatcta aatacgaccc caatgtttac agcatcaagt ataataacca attagcaact
     1381 cggactgcac aagctatttt tgatgacagc tatttgggtt attctgtggc tgtcggagat
     1441 ttcaatggtg atggcataga tgactttgtt tcaggagttc caagagcagc aaggactttg
     1501 ggaatggttt atatttatga tgggaagaac atgtcctcct tatacaattt tactggcgag
     1561 cagatggctg catatttcgg attttctgta gctgccactg acattaatgg agatgattat
     1621 gcagatgtgt ttattggagc acctctcttc atggatcgtg gctctgatgg caaactccaa
     1681 gaggtggggc aggtctcagt gtctctacag agagcttcag gagacttcca gacgacaaag
     1741 ctgaatggat ttgaggtctt tgcacggttt ggcagtgcca tagctccttt gggagatctg
     1801 gaccaggatg gtttcaatga tattgcaatt gctgctccat atgggggtga agataaaaaa
     1861 ggaattgttt atatcttcaa tggaagatca acaggcttga acgcagtccc atctcaaatc
     1921 cttgaagggc agtgggctgc tcgaagcatg ccaccaagct ttggctattc aatgaaagga
     1981 gccacagata tagacaaaaa tggatatcca gacttaattg taggagcttt tggtgtagat
     2041 cgagctatct tatacagggc cagaccagtt atcactgtaa atgctggtct tgaagtgtac
     2101 cctagcattt taaatcaaga caataaaacc tgctcactgc ctggaacagc tctcaaagtt
     2161 tcctgtttta atgttaggtt ctgcttaaag gcagatggca aaggagtact tcccaggaaa
     2221 cttaatttcc aggtggaact tcttttggat aaactcaagc aaaagggagc aattcgacga
     2281 gcactgtttc tctacagcag gtccccaagt cactccaaga acatgactat ttcaaggggg
     2341 ggactgatgc agtgtgagga attgatagcg tatctgcggg atgaatctga atttagagac
     2401 aaactcactc caattactat ttttatggaa tatcggttgg attatagaac agctgctgat
     2461 acaacaggct tgcaacccat tcttaaccag ttcacgcctg ctaacattag tcgacaggct
     2521 cacattctac ttgactgtgg tgaagacaat gtctgtaaac ccaagctgga agtttctgta
     2581 gatagtgatc aaaagaagat ctatattggg gatgacaacc ctctgacatt gattgttaag
     2641 gctcagaatc aaggagaagg tgcctacgaa gctgagctca tcgtttccat tccactgcag
     2701 gctgatttca tcggggttgt ccgaaacaat gaagccttag caagactttc ctgtgcattt
     2761 aagacagaaa accaaactcg ccaggtggta tgtgaccttg gaaacccaat gaaggctgga
     2821 actcaactct tagctggtct tcgtttcagt gtgcaccagc agtcagagat ggatacttct
     2881 gtgaaatttg acttacaaat ccaaagctca aatctatttg acaaagtaag cccagttgta
     2941 tctcacaaag ttgatcttgc tgttttagct gcagttgaga taagaggagt ctcgagtcct
     3001 gatcatatct ttcttccgat tccaaactgg gagcacaagg agaaccctga gactgaagaa
     3061 gatgttgggc cagttgttca gcacatctat gagctgagaa acaatggtcc aagttcattc
     3121 agcaaggcaa tgctccatct tcagtggcct tacaaatata ataataacac tctgttgtat
     3181 atccttcatt atgatattga tggaccaatg aactgcactt cagatatgga gatcaaccct
     3241 ttgagaatta agatctcatc tttgcaaaca actgaaaaga atgacacggt tgccgggcaa
     3301 ggtgagcggg accatctcat cactaagcgg gatcttgccc tcagtgaagg agatattcac
     3361 actttgggtt gtggagttgc tcagtgcttg aagattgtct gccaagttgg gagattagac
     3421 agaggaaaga gtgcaatctt gtacgtaaag tcattactgt ggactgagac ttttatgaat
     3481 aaagaaaatc agaatcattc ctattctctg aagtcgtctg cttcatttaa tgtcatagag
     3541 tttccttata agaatcttcc aattgaggat atcaccaact ccacattggt taccactaat
     3601 gtcacctggg gcattcagcc agcgcccatg cctgtgcctg tgtgggtgat cattttagca
     3661 gttctagcag gattgttgct actggctgtt ttggtatttg taatgtacag gatgggcttt
     3721 tttaaacggg tccggccacc tcaagaagaa caagaaaggg agcagcttca acctcatgaa
     3781 aatggtgaag gaaactcaga aactccggga tctcgagctc aagcttcgaa ttctgcagtc
     3841 gacggtaccg cgggcccggg atccccaccg gtcgccacca tggtgagcaa gggcgaggag
     3901 accacaatgg gcgtaatcaa gcccgacatg aagatcaagc tgaagatgga gggcaacgtg
     3961 aatggccacg ccttcgtgat cgagggcgag ggcgagggca agccctacga cggcaccaac
     4021 accatcaacc tggaggtgaa ggagggagcc cccctgccct tctcctacga cattctgacc
     4081 accgcgttcg cctacggcaa cagggccttc accaagtacc ccgacgacat ccccaactac
     4141 ttcaagcagt ccttccccga gggctactct tgggagcgca ccatgacctt cgaggacaag
     4201 ggcatcgtga aggtgaagtc cgacatctcc atggaggagg actccttcat ctacgagata
     4261 cacctcaagg gcgagaactt cccccccaac ggccccgtga tgcagaagaa gaccaccggc
     4321 tgggacgcct ccaccgagag gatgtacgtg cgcgacggcg tgctgaaggg cgacgtcaag
     4381 cacaagctgc tgctggaggg cggcggccac caccgcgttg acttcaagac catctacagg
     4441 gccaagaagg cggtgaagct gcccgactat cactttgtgg accaccgcat cgagatcctg
     4501 aaccacgaca aggactacaa caaggtgacc gtttacgaga gcgccgtggc ccgcaactcc
     4561 accgacggca tggacgagct gtacaagtaa gcggccgcga ctctagatca taatcagcca
     4621 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4681 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4741 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4801 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4861 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4921 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4981 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5041 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5101 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5161 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5221 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5281 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5341 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5401 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5701 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5761 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5821 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5881 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5941 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6001 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6061 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6121 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6181 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6241 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6301 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     6361 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6421 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6481 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6541 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6601 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6661 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6721 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6781 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6841 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6901 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6961 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7021 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7081 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7141 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7201 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7261 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7321 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7381 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7441 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7501 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7561 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7621 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7681 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7741 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7801 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7861 tgctcacatg ttcttt
//
