LOCUS       sequence-233121-        7608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human beta 1 ectodomain with secretion peptide, purification
            tags, and C-terminal clasp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7608)
  AUTHORS   Li J, Su Y, Xia W, Qin Y, Humphries MJ, Vestweber D, Cabanas C, Lu 
            C, Springer TA
  TITLE     Conformational equilibria and intrinsic affinities define integrin 
            activation.
  JOURNAL   EMBO J. 2017 Mar 1;36(5):629-645. doi: 10.15252/embj.201695803. Epub
            2017 Jan 25.
  PUBMED    28122868
REFERENCE   2  (bases 1 to 7608)
  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"; doi: 
            "10.15252/embj.201695803"; journalName: "EMBO J"; date: "2017-03-1- 
            1"; volume: "36"; issue: "5"; pages: "629-645"
COMMENT     Sequence Label: human b1 ecto-pIRES
FEATURES             Location/Qualifiers
     source          1..7608
                     /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      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2809..2829
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2932..2955
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     misc_feature    join(2967..3541,3542..3544,3545..3553)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3134..3151)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3361..3380
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3554..3556,3557..3559,3560..4273)
                     /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(3554..3556,3557..3559,3560..4270)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3599..3620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3860..3879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4207..4228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4396..4517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4433..4452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4487..4506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4524..4979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4661..4682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4873..4892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5006..5110
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5112..5469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5112..5303
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5135..5155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5320..5455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5504..6298
                     /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             5504..6298
                     /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(5558..5577)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6168..6187
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6486..6505)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6530..6577
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6906..7494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7395..7414
                     /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 gctagccacc
      601 atggacatgc gcgtacccgc ccagttgttg ggacttttgc tcctctggtt cagtggggtg
      661 ttggggcaga cggacgagaa tcgatgtttg aaagcgaatg caaaaagctg cggagagtgc
      721 attcaggccg gaccgaattg tggatggtgt acaaattcga catttttgca agagggaatg
      781 cccacgtcgg cccgatgtga tgacctcgag gccttgaaaa agaaaggatg tccaccggat
      841 gacatcgaga acccacgggg ctcaaaggac atcaaaaaga acaagaatgt cacaaaccgc
      901 tcgaagggaa ctgctgaaaa gcttaaaccg gaggacatca cgcagatcca gccccaacag
      961 ctggtgctcc gattgcggtc cggtgagcca cagacgttta ctctcaagtt caagagagcg
     1021 gaggattacc cgatcgatct ttactatctt atggaccttt cgtactccat gaaggatgat
     1081 ttggaaaatg tcaagagcct cggtactgac ttgatgaatg aaatgcgccg cattaccagc
     1141 gatttccgaa tcggattcgg atcatttgtg gaaaagaccg tcatgcccta catttcgacc
     1201 actccggcaa agttgcgcaa cccgtgcacc tcagaacaga actgcacctc gcctttttcg
     1261 tataagaacg tcctcagctt gacaaacaag ggtgaagtgt tcaatgagct ggtcggaaag
     1321 caaagaatct cgggaaacct cgatagcccg gagggagggt ttgatgcaat catgcaagtg
     1381 gcggtatgcg ggtcccttat tggctggaga aacgtgacca gactcctggt gttttccact
     1441 gatgcgggat ttcatttcgc cggagacggg aaactgggag ggattgtact tccgaatgac
     1501 ggacagtgtc atttggagaa caacatgtat acgatgtccc actactatga ctacccctcg
     1561 atcgcgcacc tcgtccaaaa actgagcgaa aacaatatcc agactatctt tgcggtcacg
     1621 gaggagtttc agcccgtgta taaagaactg aaaaacttga tccctaagtc cgcagtcgga
     1681 acgctgtcgg ccaacagctc gaatgtgatc cagttgatca tcgatgcgta taactcactt
     1741 tcatcagagg tcatccttga gaacggaaag cttagcgaag gagtgacaat ctcgtacaaa
     1801 tcatattgca agaatggagt gaatggcacc ggcgagaatg gccggaaatg ttcaaacatt
     1861 tccatcggcg acgaggtcca gtttgagatc agcattactt ccaataagtg ccctaagaaa
     1921 gactcggact cattcaaaat tcggcctctc gggtttaccg aagaggtaga ggtgatcctc
     1981 cagtacattt gtgaatgcga gtgtcaatca gagggtatcc ccgaaagccc caagtgtcat
     2041 gagggtaacg gaacgttcga atgcggggca tgtagatgta acgaaggtcg cgtcggaagg
     2101 cactgcgaat gctcgactga tgaggtaaac tcggaagaca tggatgcgta ttgccgaaaa
     2161 gagaattcga gcgaaatctg ctcaaacaac ggggaatgtg tgtgcggaca atgtgtctgc
     2221 cggaaacggg ataacacaaa tgagatctac tccggaaagt tttgcgaatg cgacaatttc
     2281 aactgtgaca ggtccaacgg tcttatctgt ggtggtaatg gagtctgcaa gtgccgcgtc
     2341 tgcgagtgta atccgaatta cacaggctca gcgtgtgatt gcagcctcga cacgtcaaca
     2401 tgcgaggcgt ccaacgggca gatttgtaac gggcgaggga tttgcgagtg tggggtgtgc
     2461 aaatgcactg accccaaatt ccaggggcag acgtgcgaga tgtgccagac gtgtcttggt
     2521 gtctgtgcgg agcataagga atgcgtgcag tgcagagcgt tcaacaaagg ggaaaagaaa
     2581 gatacgtgta cgcaagaatg ttcgtatttc aatatcacga aggtggagtc gagggataag
     2641 ctcccgcagc cagtacagcc ggaccccgtc agccattgca aggaaaagga cgtggatgat
     2701 tgctggttct atttcacata ctccgtgaat ggtaataatg aggtaatggt acatgtcgtg
     2761 gagaaccccg aatgccccac gggtcccgac gggcaccaac accaactcga gaatctctac
     2821 ttccaaggtg gggaaggccc agcccagtgt aagaagaaac tccaggcgct gaagaaaaag
     2881 aacgcgcagt tgaaatggaa gctccaagca ctgaaaaaga aattggcgca gcatcaccac
     2941 catcaccatc atcattaaag gatccgcccc tctccctccc ccccccctaa cgttactggc
     3001 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     3061 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     3121 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     3181 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     3241 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     3301 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3361 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3421 atgggatctg atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa
     3481 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     3541 tatggccaca accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     3781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     4021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     4081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4141 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     4201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     4261 gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     4321 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4381 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4441 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4501 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     4561 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4621 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4681 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4741 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4801 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4861 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4921 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4981 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5041 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5101 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     5161 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5221 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5281 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5341 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5401 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5461 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5521 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5581 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5641 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     5701 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5761 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5821 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5881 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5941 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6001 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     6061 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6121 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6181 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6241 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6301 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6361 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6421 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     6481 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     6541 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     6601 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     6661 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     6721 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     6781 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6841 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6901 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6961 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7021 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7081 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7141 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7201 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7261 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7321 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7381 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7441 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7501 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7561 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
