LOCUS       sequence-350818-        7069 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 7069)
  AUTHORS   Tsuruta D, Gonzales M, Hopkinson SB, Otey C, Khuon S, Goldman RD, 
            Jones JC
  TITLE     Microfilament-dependent movement of the beta3 integrin subunit 
            within focal contacts of endothelial cells.
  JOURNAL   FASEB J. 2002 Jun . 16(8):866-8.
  PUBMED    11967230
REFERENCE   2  (bases 1 to 7069)
  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: "FASEB J. 
            2002 Jun . 16(8):866-8."
COMMENT     Sequence Label: beta-3-integrin-YFP
FEATURES             Location/Qualifiers
     source          1..7069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    340..461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(377..396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     431..450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(468..923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(605..626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     817..836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        950..1054
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1056..1413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1056..1247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1079..1099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1264..1399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1448..2242
                     /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             1448..2242
                     /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(1502..1521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2112..2131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2430..2449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2474..2521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2850..3438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3339..3358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3613..3916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3917..4120
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4071..4091
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6561..6570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6567..6569,6570..6572,6573..7069,1..217)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6612..6633)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6873..6892)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
       61 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
      121 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      181 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
      241 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      301 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      361 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
      421 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
      481 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
      541 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
      601 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
      661 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
      721 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
      781 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
      841 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
      901 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      961 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1021 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     1081 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1141 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     1201 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     1261 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1321 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     1381 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     1441 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     1501 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     1561 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     1621 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     1681 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     1741 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     1801 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     1861 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     1921 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     1981 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     2041 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     2101 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2161 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2221 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2281 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2341 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2401 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     2461 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     2521 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2581 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2641 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     2701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3541 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     3601 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3661 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3721 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3781 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3841 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3901 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3961 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4021 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4081 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     4141 ccgctagcgc taccggactc agatctcgag ctcaagctta cgagatgcga gcgcggccgc
     4201 ggccccggcc gctctgggcg actgtgctgg cgctgggggc gctggcgggc gttggcgtag
     4261 gagggcccaa catctgtacc acgcgaggtg tgagctcctg ccagcagtgc ctggctgtga
     4321 gccccatgtg tgcctggtgc tctgatgagg ccctgcctct gggctcacct cgctgtgacc
     4381 tgaaggagaa tctgctgaag gataactgtg ccccagaatc catcgagttc ccagtgagtg
     4441 aggcccgagt actagaggac aggcccctca gcgacaaggg ctccggagac agctcccagg
     4501 tcactcaagt cagtccccag aggattgcac tccggctccg gccagatgat tcgaagaatt
     4561 tctccatcca agtgcggcag gtggaggatt accctgtgga catctactac ttgatggacc
     4621 tgtcttactc catgaaggat gatctgtgga gcatccagaa cctgggtacc aagctggcca
     4681 cccagatgcg aaagctcacc agtaacctgc ggattggctt cggggcattt gtggacaagc
     4741 ctgtgtcacc atacatgtat atctccccac cagaggccct cgaaaacccc tgctatgata
     4801 tgaagaccac ctgcttgccc atgtttggct acaaacacgt gctgacgcta actgaccagg
     4861 tgacccgctt caatgaggaa gtgaagaagc agagtgtgtc acggaaccga gatgccccag
     4921 agggtggctt tgatgccatc atgcaggcta cagtctgtga tgaaaagatt ggctggagga
     4981 atgatgcatc ccacttgctg gtgtttacca ctgatgccaa gactcatata gcattggacg
     5041 gaaggctggc aggcatcgtc cagcctaatg acgggcagtg tcatgttggt agtgacaatc
     5101 attactctgc ctccactacc atggattatc cctctttggg gctgatgact gagaagctat
     5161 cccagaaaaa catcaatttg atctttgcag tgactgaaaa tgtagtcaat ctctatcaga
     5221 actatagtga gctcatccca gggaccacag ttggggttct gtccatggat tccagcaatg
     5281 tcctccagct cattgttgat gcttatggga aaatccgttc taaagtcgag ctggaagtgc
     5341 gtgacctccc tgaagagttg tctctatcct tcaatgccac ctgcctcaac aatgaggtca
     5401 tccctggcct caagtcttgt atgggactca agattggaga cacggtgagc ttcagcattg
     5461 aggccaaggt gcgaggctgt ccccaggaga aggagaagtc ctttaccata aagcccgtgg
     5521 gcttcaagga cagcctgatc gtccaggtca cctttgattg tgactgtgcc tgccaggccc
     5581 aagctgaacc taatagccat cgctgcaaca atggcaatgg gacctttgag tgtggggtat
     5641 gccgttgtgg gcctggctgg ctgggatccc agtgtgagtg ctcagaggag gactatcgcc
     5701 cttcccagca ggacgagtgc agcccccgag agggtcagcc cgtctgcagc cagcggggcg
     5761 agtgcctctg tggtcaatgt gtctgccaca gcagcgactt tggcaagatc acgggcaagt
     5821 actgcgagtg tgacgacttc tcctgtgtcc gctacaaggg ggagatgtgc tcaggccatg
     5881 gccagtgcag ctgtggggac tgcctgtgtg actccgactg gaccggctac tactgcaact
     5941 gtaccacgcg tactgacacc tgcatgtcca gcaatgggct gctgtgcagc ggccgcggca
     6001 aatgtgaatg tggcagctgt gtctgtatcc agccgggctc ctatggggac acctgtgaga
     6061 agtgccccac ctgcccagat gcctgcacct ttaagaaaga atgtgtggag tgtaagaagt
     6121 ttgaccgggg agccctacat gacgaaaata cctgcaaccg ttactgccgt gacgagattg
     6181 agtcagtgaa agagcttaag gacactggca aggatgcagt gaattgtacc tataagaatg
     6241 aggatgactg tgtcgtcaga ttccagtact atgaagattc tagtggaaag tccatcctgt
     6301 atgtggtaga agagccagag tgtcccaagg gccctgacat cctggtggtc ctgctctcag
     6361 tgatgggggc cattctgctc attggccttg ccgccctgct catctggaaa ctcctcatca
     6421 ccatccacga ccgaaaagaa ttcgctaaat ttgaggaaga acgcgccaga gcaaaatggg
     6481 acacagccaa caacccactg tataaagagg ccacgtctac cttcaccaat atcacgtacc
     6541 ggggcacaga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     6601 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     6661 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     6721 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct
     6781 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     6841 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     6901 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     6961 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     7021 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagat
//
