LOCUS       sequence-7711-ex        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7113)
  TITLE     Other integrin beta6 plasmids
REFERENCE   2  (bases 1 to 7113)
  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: integrin beta6 EGFP-N3
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3053..3062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3059..3061,3062..3064,3065..3778)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3059..3061,3062..3064,3065..3775)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3104..3125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3365..3384)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3712..3733
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3901..4022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3938..3957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3992..4011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4029..4484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4166..4187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4378..4397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4511..4615
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4617..4974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4617..4808
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4640..4660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4825..4960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5009..5803
                     /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             5009..5803
                     /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(5063..5082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5673..5692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5991..6010)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6035..6082
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6411..6999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6900..6919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat ccattgtgct ctaaaggcaa gaactgaaac gaatggggat
      661 tgaactgctt tgcctgttct ttctatttct aggaaggaat gatcacgtac aaggtggctg
      721 tgccctggga ggtgcagaaa cctgtgaaga ctgcctgctt attggacctc agtgtgcctg
      781 gtgtgctcag gagaatttta ctcatccatc tggagttggc gaaaggtgtg ataccccagc
      841 aaacctttta gctaaaggat gtcaattaaa cttcatcgaa aaccctgtct cccaagtaga
      901 aatacttaaa aataagcctc tcagtgtagg cagacagaaa aatagttctg acattgttca
      961 gattgcgcct caaagcttga tccttaagtt gagaccaggt ggtgcgcaga ctctgcaggt
     1021 gcatgtccgc cagactgagg actacccggt ggatttgtat tacctcatgg acctctccgc
     1081 ctccatggat gacgacctca acacaataaa ggagctgggc tcccggcttt ccaaagagat
     1141 gtctaaatta accagcaact ttagactggg cttcggatct tttgtggaaa aacctgtatc
     1201 ccctttcgtg aaaacaacac cagaagaaat tgccaaccct tgcagtagta ttccatactt
     1261 ctgtttacct acatttggat tcaagcacat tttgccattg acaaatgatg ctgaaagatt
     1321 caatgaaatt gtgaagaatc agaaaatttc tgctaatatt gacacacccg aaggtggatt
     1381 tgatgcaatt atgcaagctg ctgtgtgtaa ggaaaaaatt ggctggcgga atgactccct
     1441 ccacctcctg gtctttgtga gtgatgctga ttctcatttt ggaatggaca gcaaactagc
     1501 aggcatcgtc attcctaatg acgggctctg tcacttggac agcaagaatg aatactccat
     1561 gtcaactgtc ttggaatatc caacaattgg acaactcatt gataaactgg tacaaaacaa
     1621 cgtgttattg atcttcgctg taacccaaga acaagttcat ttatatgaga attacgcaaa
     1681 acttattcct ggagctacag taggtctact tcagaaggac tccggaaaca ttctccagct
     1741 gatcatctca gcttatgaag aactgcggtc tgaggtggaa ctggaagtat taggagacac
     1801 tgaaggactc aacttgtcat ttacagccat ctgtaacaac ggtaccctct tccaacacca
     1861 aaagaaatgc tctcacatga aagtgggaga cacagcttcc ttcagcgtga ctgtgaatat
     1921 cccacactgc gagagaagaa gcaggcacat tatcataaag cctgtggggc tgggggatgc
     1981 cctggaatta cttgtcagcc cagaatgcaa ctgcgactgt cagaaagaag tggaagtgaa
     2041 cagctccaaa tgtcaccacg ggaacggctc tttccagtgt ggggtgtgtg cctgccaccc
     2101 tggccacatg gggcctcgct gtgagtgtgg cgaggacatg ctgagcacag attcctgcaa
     2161 ggaggcccca gatcatccct cctgcagcgg aaggggtgac tgctactgtg ggcagtgtat
     2221 ctgccacttg tctccctatg gaaacattta tgggccttat tgccagtgtg acaatttctc
     2281 ctgcgtgaga cacaaagggc tgctctgcgg aggtaacggc gactgtgact gtggtgaatg
     2341 tgtgtgcagg agcggctgga ctggcgagta ctgcaactgc accaccagca cggactcctg
     2401 cgtctctgaa gatggagtgc tctgcagcgg gcgcggggac tgtgtttgtg gcaagtgtgt
     2461 ttgcacaaac cctggagcct caggaccaac ctgtgaacga tgtcctacct gtggtgaccc
     2521 ctgtaactct aaacggagct gcattgagtg ccacctgtca gcagctggcc aagcccgaga
     2581 agaatgtgtg gacaagtgca aactagctgg tgcgaccatc agtgaagaag aagatttctc
     2641 aaaggatggt tctgtttcct gctctctgca aggagaaaat gaatgtctta ttacattcct
     2701 aataactaca gataatgagg ggaaaaccat cattcacagc atcaatgaaa aagattgtcc
     2761 gaagcctcca aacattccca tgatcatgtt aggggtttcc ctggctattc ttctcatcgg
     2821 ggttgtccta ctgtgcatct ggaagctact ggtgtcattt catgatcgta aagaagttgc
     2881 caaatttgaa gcagaacgat caaaagccaa gtggcaaacg ggaaccaatc cactctacag
     2941 aggatccaca agtactttta aaaatgtaac ttataaacac agggaaaaac aaaaggtaga
     3001 cctttccaca gattgctcta gagcggccgc caccgcgggc ccgggatcca tcgccaccat
     3061 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3121 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3181 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3241 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3301 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3361 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3421 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3481 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3541 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3601 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3661 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3721 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     3781 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3841 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3961 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4021 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4081 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4141 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4201 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4261 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4321 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4381 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4441 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     4501 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4561 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     4621 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4741 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4801 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4861 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4921 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4981 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5041 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5101 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5161 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     5221 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5281 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5341 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5401 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5461 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5521 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5581 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5641 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5701 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5761 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5821 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5881 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5941 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     6001 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     6061 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     6121 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     6181 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     6241 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     6301 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6361 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6421 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6481 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6541 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     6601 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6661 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6721 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6781 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6841 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6901 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6961 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7021 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7081 gattctgtgg ataaccgtat taccgccatg cat
//
