LOCUS       Exported                6997 bp ds-DNA     circular SYN 28-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    EBFP2-Kindlin2-3KA [Homo Sapiens]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6997)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6997
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1310..1319
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1316..2032
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     misc_feature    2207..4246
                     /label=Kindlin2-3KA [Homo Sapiens]
     polyA_signal    4488..4609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4616..5071)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5098..5202
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5204..5561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5204..5395
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      5412..5547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5596..6390
                     /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             5596..6390
                     /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"
     polyA_signal    6622..6669
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgagggg cgagggcgag ggcgatgcca ccaacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg agccacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctagtgaa
     1681 ccgcatcgag ctgaagggcg tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac ttcaacagcc acaacatcta tatcatggcc gtcaagcaga agaacggcat
     1801 caaggtgaac ttcaagatcc gccacaacgt ggaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca gccactacct
     1921 gagcacccag tccgtgctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttccgc accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     2041 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc ctcgacccac
     2101 gcgtccggcg gcgcgaggct tcgctttcct ccagagcagc gacgcgggtg ccgccagcct
     2161 cgcctgttgc cgcgggagcc cggcgggagg agcgcggaag ggagccatgg ctctggacgg
     2221 gataaggatg ccagatggct gctacgcgga cgggacgtgg gaactgagtg tccatgtgac
     2281 ggacctgaac cgcgatgtca ccctgagagt gaccggcgag gtgcacattg gaggcgtgat
     2341 gcttaagctg gtggagaaac tcgatgtaaa aaaagattgg tctgaccatg ctctctggtg
     2401 ggaaaagaag agaacttggc ttctgaagac acattggacc ttagataagt atggtattca
     2461 ggcagatgct aagcttcagt tcacccctca gcacaaactg ctccgcctgc agcttcccaa
     2521 catgaagtat gtgaaggtga aagtgaattt ctctgataga gtcttcaaag ctgtttctga
     2581 catctgtaag acttttaata tcagacaccc cgaagaactt tctctcttaa agaaacccag
     2641 agatccaaca aagaaaaaaa agaagaagct agatgaccag tctgaagatg aggcacttga
     2701 attagagggg cctcttatca ctcctggatc aggaagtata tattcaagcc caggactgta
     2761 tagtaaaaca atgaccccca cttatgatgc tcatgatgga agccccttgt caccaacttc
     2821 tgcttggttt ggtgacagtg ctttgtcaga aggcaatcct ggtatacttg ctgtcagtca
     2881 accaatcacg tcaccagaaa tcttggcaaa aatgttcaag cctcaagctc ttcttgataa
     2941 agcaaaaatc aaccaaggat ggcttgattc ctcaagatct ctcatggaac aagatgtgaa
     3001 ggaaaatgag gccttgctgc tccgattcaa gtattacagc ttttttgatt tgaatccaaa
     3061 gtatgatgca atcagaatca atcagcttta tgagcaggcc aaatgggcca ttctcctgga
     3121 agagattgaa tgcacagaag aagaaatgat gatgtttgca gccctgcagt atcatatcaa
     3181 taagctgtca atcatgacat cagagaatca tttgaacaac agtgacaaag aagttgatga
     3241 agttgatgct gccctttcag acctggagat tactctggaa gggggtaaaa cgtcaacaat
     3301 tttgggtgac attacttcca ttcctgaact tgctgactac attaaagttt tcaagccagc
     3361 ggcgctgact ctgaaaggtt acgcgcaata ttggtgcacc ttcaaagaca catccatttc
     3421 ttgttataag agcaaagaag aatccagtgg cacaccagct catcagatga acctcagggg
     3481 atgtgaagtt accccagatg taaacatttc aggccaaaaa tttaacatta aactcctgat
     3541 tccagttgca gaaggcatga atgaaatctg gcttcgttgt gacaatgaaa aacagtatgc
     3601 acactggatg gcagcctgca gattagcctc caaaggcaag accatggcgg acagttctta
     3661 caacttagaa gttcagaata ttctttcctt tctgaagatg cagcatttaa acccagatcc
     3721 tcagttaata ccagagcaga tcacgactga tataactcct gaatgtttgg tgtctccccg
     3781 ctatctaaaa aagtataaga acaagcagat aacagcgaga atcttggagg cccatcagaa
     3841 tgtagctcag atgagtctaa ttgaagccaa gatgagattt attcaagctt ggcagtcact
     3901 acctgaattt ggcatcactc acttcattgc aaggttccaa gggggcaaaa aagaagaact
     3961 tattggaatt gcatacaaca gactgattcg gatggatgcc agcactggag atgcaattaa
     4021 aacatggcgt ttcagcaaca tgaaacagtg gaatgtcaac tgggaaatca aaatggtcac
     4081 cgtagagttt gcagatgaag tacgattgtc cttcatttgt actgaagtag attgcaaagt
     4141 ggttcatgaa ttcattggtg gctacatatt tctctcaaca cgtgcaaaag accaaaacga
     4201 gagtttagat gaagagatgt tctacaaact taccagtggt tgggtgtgaa taggaaaact
     4261 tgtaataaaa ctccacagcc ataacaatat ttaactttaa agctattgtt tcttatatgc
     4321 tgcttaataa agtaagcttg aactttaaaa aaaaaaaaaa gggcggccgc tctagataac
     4381 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4441 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4501 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4561 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     4621 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4681 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4741 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4801 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4861 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4921 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4981 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5041 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     5101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     5221 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5281 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5341 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5401 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5461 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5521 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     5581 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5641 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5701 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5761 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5821 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5881 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5941 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6001 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6061 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6121 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6181 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6241 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6301 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6361 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6421 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6481 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6541 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     6601 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     6661 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     6721 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     6781 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     6841 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6901 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6961 gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
