LOCUS       sequence-366177-        7127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mPA-GFP tagged Integrin beta-1 in mammalian cells on a CMV
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7127)
  AUTHORS   Jain K, Kanchanawong P, Sheetz MP, Zhou X, Cai H, Changede R
  TITLE     Ligand functionalization of titanium nanopattern enables the 
            analysis of cell-ligand interactions by super-resolution microscopy.
  JOURNAL   Nat Protoc. 2022 Jul 27. pii: 10.1038/s41596-022-00717-3. doi: 
            10.1038/s41596-022-00717-3.
  PUBMED    35896742
REFERENCE   2  (bases 1 to 7127)
  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: "Nat Protoc.
            2022 Jul 27. pii: 10.1038/s41596-022-00717-3. doi: 
            10.1038/s41596-022-00717-3."
COMMENT     Sequence Label: Integrin Beta1-mPA-GFP
FEATURES             Location/Qualifiers
     source          1..7127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        375..578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     529..549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3077..3086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3083..3085,3086..3088,3089..3799)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3128..3149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3389..3408)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3736..3757
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3925..4046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3962..3981)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4016..4035
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4053..4508)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4190..4211)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4402..4421
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4535..4639
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4641..4998
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4641..4832
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4664..4684)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4849..4984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5033..5827
                     /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             5033..5827
                     /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(5087..5106)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5697..5716
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6015..6034)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6059..6106
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6435..7023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6924..6943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      601 gctagcgcta ccggactcag atctcgagac cgccatgaat ttacaaccaa ttttctggat
      661 tggactgatc agttcagttt gctgtgtgtt tgctcaaaca gatgaaaata gatgtttaaa
      721 agcaaatgcc aaatcatgtg gagaatgtat acaagcaggg ccaaattgtg ggtggtgcac
      781 aaattcaaca tttttacagg aaggaatgcc tacttctgca cgatgtgatg atttagaagc
      841 cttaaaaaag aagggttgcc ctccagatga catagaaaat cccagaggct ccaaagatat
      901 aaagaaaaat aaaaatgtaa ccaaccgtag caaaggaaca gcagagaagc tcaagccaga
      961 ggatattact cagatccaac cacagcagtt ggttttgcga ttaagatcag gggagccaca
     1021 gacatttaca ttaaaattca agagagctga agactatccc attgacctct actaccttat
     1081 ggacctgtct tactcaatga aagacgattt ggagaatgta aaaagtcttg gaacagatct
     1141 gatgaatgaa atgaggagga ttacttcgga cttcagaatt ggatttggct catttgtgga
     1201 aaagactgtg atgccttaca ttagcacaac accagctaag ctcaggaacc cttgcacaag
     1261 tgaacagaac tgcaccagcc catttagcta caaaaatgtg ctcagtctta ctaataaagg
     1321 agaagtattt aatgaacttg ttggaaaaca gcgcatatct ggaaatttgg attctccaga
     1381 aggtggtttc gatgccatca tgcaagttgc agtttgtgga tcactgattg gctggaggaa
     1441 tgttacacgg ctgctggtgt tttccacaga tgccgggttt cactttgctg gagatgggaa
     1501 acttggtggc attgttttac caaatgatgg acaatgtcac ctggaaaata atatgtacac
     1561 aatgagccat tattatgatt atccttctat tgctcacctt gtccagaaac tgagtgaaaa
     1621 taatattcag acaatttttg cagttactga agaatttcag cctgtttaca aggagctgaa
     1681 aaacttgatc cctaagtcag cagtaggaac attatctgca aattctagca atgtaattca
     1741 gttgatcatt gatgcataca attccctttc ctcagaagtc attttggaaa acggcaaatt
     1801 gtcagaagga gtaacaataa gttacaaatc ttactgcaag aacggggtga atggaacagg
     1861 ggaaaatgga agaaaatgtt ccaatatttc cattggagat gaggttcaat ttgaaattag
     1921 cataacttca aataagtgtc caaaaaagga ttctgacagc tttaaaatta ggcctctggg
     1981 ctttacggag gaagtagagg ttattcttca gtacatctgt gaatgtgaat gccaaagcga
     2041 aggcatccct gaaagtccca agtgtcatga aggaaatggg acatttgagt gtggcgcgtg
     2101 caggtgcaat gaagggcgtg ttggtagaca ttgtgaatgc agcacagatg aagttaacag
     2161 tgaagacatg gatgcttact gcaggaaaga aaacagttca gaaatctgca gtaacaatgg
     2221 agagtgcgtc tgcggacagt gtgtttgtag gaagagggat aatacaaatg aaatttattc
     2281 tggcaaattc tgcgagtgtg ataatttcaa ctgtgataga tccaatggct taatttgtgg
     2341 aggaaatggt gtttgcaagt gtcgtgtgtg tgagtgcaac cccaactaca ctggcagtgc
     2401 atgtgactgt tctttggata ctagtacttg tgaagccagc aacggacaga tctgcaatgg
     2461 ccggggcatc tgcgagtgtg gtgtctgtaa gtgtacagat ccgaagtttc aagggcaaac
     2521 gtgtgagatg tgtcagacct gccttggtgt ctgtgctgag cataaagaat gtgttcagtg
     2581 cagagccttc aataaaggag aaaagaaaga cacatgcaca caggaatgtt cctattttaa
     2641 cattaccaag gtagaaagtc gggacaaatt accccagccg gtccaacctg atcctgtgtc
     2701 ccattgtaag gagaaggatg ttgacgactg ttggttctat tttacgtatt cagtgaatgg
     2761 gaacaacgag gtcatggttc atgttgtgga gaatccagag tgtcccactg gtccagacat
     2821 cattccaatt gtagctggtg tggttgctgg aattgttctt attggccttg cattactgct
     2881 gatatggaag cttttaatga taattcatga cagaagggag tttgctaaat ttgaaaagga
     2941 gaaaatgaat gccaaatggg acacgggtga aaatcctatt tataagagtg ccgtaacaac
     3001 tgtggtcaat ccgaagtatg agggaaaagg tagcgcatcc gggggtgcgt caggcggcag
     3061 cggatctgga ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     3121 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3181 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3241 gctgcccgtg ccctggccca ccctcgtgac caccttcagc tacggcgtgc agtgcttcag
     3301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3361 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3481 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3541 catggccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     3601 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3661 cgtgctgctg cccgacaacc actacctgag ccaccagtcc aagctgagca aagaccccaa
     3721 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3781 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     3841 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3901 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3961 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4021 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     4081 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4141 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4201 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4261 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4321 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4381 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4441 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4501 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4621 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4681 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4741 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4801 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4861 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4921 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4981 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5041 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5101 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5161 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5221 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5281 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5341 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5401 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5461 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5521 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5581 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5641 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5701 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5761 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5821 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5881 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5941 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6001 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6061 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6121 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6181 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6241 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6541 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattac
//
