LOCUS       Exported                6988 bp ds-DNA     circular SYN 21-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    EBFP2-Kindlin1 [Homo Sapiens]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6988)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6988
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        765..1068
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1069..1272
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1311..1320
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1317..2033
                     /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    2208..4238
                     /label=Kindlin1 [Homo Sapiens]
     polyA_signal    4480..4601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4608..5063)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5090..5194
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5196..5553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5196..5387
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      5404..5539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5588..6382
                     /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             5588..6382
                     /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    6614..6661
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
      721 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      781 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      841 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      901 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      961 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1021 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1081 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1141 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1201 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1261 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg
     1321 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1381 acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca
     1441 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1501 tgaccaccct gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1561 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1621 aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctagtga
     1681 accgcatcga gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc
     1741 tggagtacaa cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca
     1801 tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc
     1861 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc
     1921 tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1981 tggagttccg caccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac
     2041 tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc cctcgaccca
     2101 cgcgtccggc ggcgcgaggc ttcgctttcc tccagagcag cgacgcgggt gccgccagcc
     2161 tcgcctgttg ccgcgggagc ccggcgggag gagcgcggaa gggagccatg ctgtcatcca
     2221 ctgactttac atttgcttcc tgggagcttg tggtccgcgt tgaccatccc aatgaagagc
     2281 agcagaaaga cgtcacactg agagtatctg gagaccttca cgttggagga gtgatgctca
     2341 agttagtaga acagatcaat atatcccaag actggtcaga ctttgctctt tggtgggaac
     2401 agaagcattg ctggcttctg aaaacccact ggaccctgga caaatatggg gtccaggcag
     2461 atgcaaagct tctcttcacc cctcagcata aaatgctgcg ccttcgtctg ccgaatttga
     2521 agatggtgag gttgcgagtc agcttctcag ctgtggtttt taaagctgtc agtgatatct
     2581 gcaaaatcct gaatattaga agatcagaag agctttcctt gttaaagccg tctggtgact
     2641 attttaagaa gaagaagaaa aaagacaaaa ataataagga acccataatt gaagatattc
     2701 taaacctgga gagttctcca acagcttcag gttcatcagt aagtcctggt ttatacagta
     2761 aaaccatgac ccctatatat gaccccatca atggaacacc agcatcatcc accatgactt
     2821 ggttcagtga cagccctttg acggaacaaa actgcagcat cctcgcattc agccaacccc
     2881 cccagtcccc agaagcactt gcggatatgt accagcctcg gtctctggtt gataaagcca
     2941 agctcaatgc aggttggcta gactcctcac gctcccttat ggaacaaggc atccaagagg
     3001 atgagcagct gctcttacga tttaaatatt attctttctt cgacttgaat cctaaatatg
     3061 atgctgtccg aataaaccaa ctctatgagc aagccaggtg ggccattctc ttagaagaaa
     3121 ttgattgcac agaggaagaa atgttgatct ttgcagctct acagtaccac attagcaaac
     3181 tgtcgttgtc tgctgaaaca caggattttg caggcgagtc cgaggttgat gaaatagaag
     3241 cggcgctttc taatttggaa gtaaccctag aaggtggaaa agcggacagc cttttggagg
     3301 acattactga tatccctaaa cttgcagata atctcaaatt atttaggccc aagaagttac
     3361 taccaaaagc tttcaaacaa tattggttta tctttaaaga cacatccata gcatacttta
     3421 aaaataagga acttgaacaa ggagaaccac tagaaaaact aaatcttaga ggctgcgaag
     3481 ttgtgcccga tgtaaatgta gcaggaagaa aatttggaat caagttacta atccctgttg
     3541 ccgatggtat gaatgaaatg tatttgagat gtgaccatga gaatcaatac gcccaatgga
     3601 tggctgcctg catgttggca tcgaagggca aaaccatggc agacagctcc taccagccag
     3661 aggtcctcaa catcctttca tttctgagga tgaaaaacag gaactctgca tctcaggtgg
     3721 cttccagtct cgaaaacatg gatatgaacc cagaatgttt tgtgtcacca cggtgtgcaa
     3781 aaagacacaa atccaaacag ctggccgccc ggatcctgga ggcgcaccag aacgtggccc
     3841 agatgcccct ggtcgaagcc aagctgcggt tcatccaggc gtggcagtca ctgcctgagt
     3901 ttggcctcac ctactacctt gtcagattta aaggaagcaa aaaagatgac attctgggag
     3961 tttcatataa caggttgatt aaaattgatg cagccaccgg gattccagtg acaacatgga
     4021 gattcacaaa tatcaaacag tggaatgtaa actgggaaac ccggcaggtg gtcatcgagt
     4081 ttgaccaaaa cgtctttact gctttcacct gcctgagtgc agattgcaag attgtgcacg
     4141 agtacattgg cggctacatt ttcttgtcca cccgctccaa ggaccagaat gaaacactcg
     4201 atgaggactt gttccacaaa ttgaccggcg gtcaggattg aataggaaaa cttgtaataa
     4261 aactccacag ccataacaat atttaacttt aaagctattg tttcttatat gctgcttaat
     4321 aaagtaagct tgaactttaa aaaaaaaaaa aagggcggcc gctctagata actgatcata
     4381 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4441 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4501 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4561 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     4621 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4681 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4741 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4801 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4861 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4921 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4981 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5041 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5101 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5161 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     5221 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5281 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5341 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5401 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5461 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5521 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     5581 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5641 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5701 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5761 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5821 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5881 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5941 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6001 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6061 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6121 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6181 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6241 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6301 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6361 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6421 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6481 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6541 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     6601 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6661 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6721 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     6781 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     6841 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6901 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6961 gaccccgtag aaaagatcaa aggatctt
//
