LOCUS       sequence-151351-        7013 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 7013)
  AUTHORS   Kim M, Carman CV, Springer TA
  TITLE     Bidirectional transmembrane signaling by cytoplasmic domain 
            separation in integrins.
  JOURNAL   Science 2003 Sep 19;301(5640):1720-5.
  PUBMED    14500982
REFERENCE   2  (bases 1 to 7013)
  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: "Science"; 
            date: "2003-09-19"; volume: "301(5640)"; pages: "1720-5"
COMMENT     Sequence Label: Integrin beta 2 - mYFP
FEATURES             Location/Qualifiers
     source          1..7013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2908..2917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2914..2916,2917..2919,2920..3633)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2959..2980)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3220..3239)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3567..3588
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3756..3877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3793..3812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3847..3866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3884..4339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4021..4042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4233..4252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4366..4470
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4472..4829
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4472..4663
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4495..4515)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4680..4815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4864..5658
                     /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             4864..5658
                     /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(4918..4937)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5528..5547
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5846..5865)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5890..5937
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6266..6854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6755..6774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttgccaccat gctgggcctg
      601 cgccccccac tgctcgccct ggtggggctg ctctccctcg ggtgcgtcct ctctcaggag
      661 tgcacgaagt tcaaggtcag cagctgccgg gaatgcatcg agtcggggcc cggctgcacc
      721 tggtgccaga agctgaactt cacagggccg ggggatcctg actccattcg ctgcgacacc
      781 cggccacagc tgctcatgag gggctgtgcg gctgacgaca tcatggaccc cacaagcctc
      841 gctgaaaccc aggaagacca caatgggggc cagaagcagc tgtccccaca aaaagtgacg
      901 ctttacctgc gaccaggcca ggcagcagcg ttcaacgtga ccttccggcg ggccaagggc
      961 taccccatcg acctgtacta tctgatggac ctctcctact ccatgcttga tgacctcagg
     1021 aatgtcaaga agctaggtgg cgacctgctc cgggccctca acgagatcac cgagtccggc
     1081 cgcattggct tcgggtcctt cgtggacaag accgtgctgc cgttcgtgaa cacgcaccct
     1141 gataagctgc gaaacccatg ccccaacaag gagaaagagt gccagccccc gtttgccttc
     1201 aggcacgtgc tgaagctgac caacaactcc aaccagtttc agaccgaggt cgggaagcag
     1261 ctgatttccg gaaacctgga tgcacccgag ggtgggctgg acgccatgat gcaggtcgcc
     1321 gcctgcccgg aggaaatcgg ctggcgcaac gtcacgcggc tgctggtgtt tgccactgat
     1381 gacggcttcc atttcgcggg cgacggaaag ctgggcgcca tcctgacccc caacgacggc
     1441 cgctgtcacc tggaggacaa cttgtacaag aggagcaacg aattcgacta cccatcggtg
     1501 ggccagctgg cgcacaagct ggctgaaaac aacatccagc ccatcttcgc ggtgaccagt
     1561 aggatggtga agacctacga gaaactcacc gagatcatcc ccaagtcagc cgtgggggag
     1621 ctgtctgagg actccagcaa tgtggtccat ctcattaaga atgcttacaa taaactctcc
     1681 tccagggtct tcctggatca caacgccctc cccgacaccc tgaaagtcac ctacgactcc
     1741 ttctgcagca atggagtgac gcacaggaac cagcccagag gtgactgtga tggcgtgcag
     1801 atcaatgtcc cgatcacctt ccaggtgaag gtcacggcca cagagtgcat ccaggagcag
     1861 tcgtttgtca tccgggcgct gggcttcacg gacatagtga ccgtgcaggt tcttccccag
     1921 tgtgagtgcc ggtgccggga ccagagcaga gaccgcagcc tctgccatgg caagggcttc
     1981 ttggagtgcg gcatctgcag gtgtgacact ggctacattg ggaaaaactg tgagtgccag
     2041 acacagggcc ggagcagcca ggagctggaa ggaagctgcc ggaaggacaa caactccatc
     2101 atctgctcag ggctggggga ctgtgtctgc gggcagtgcc tgtgccacac cagcgacgtc
     2161 cccggcaagc tgatatacgg gcagtactgc gagtgtgaca ccatcaactg tgagcgctac
     2221 aacggccagg tctgcggcgg cccggggagg gggctctgct tctgcgggaa gtgccgctgc
     2281 cacccgggct ttgagggctc agcgtgccag tgcgagagga ccactgaggg ctgcctgaac
     2341 ccgcggcgtg ttgagtgtag tggtcgtggc cggtgccgct gcaacgtatg cgagtgccat
     2401 tcaggctacc agctgcctct gtgccaggag tgccccggct gcccctcacc ctgtggcaag
     2461 tacatctcct gcgccgagtg cctgaagttc gaaaagggcc cctttgggaa gaactgcagc
     2521 gcggcgtgtc cgggcctgca gctgtcgaac aaccccgtga agggcaggac ctgcaaggag
     2581 agggactcag agggctgctg ggtggcctac acgctggagc agcaggacgg gatggaccgc
     2641 tacctcatct atgtggatga gagccgagag tgtgtggcag gccccaacat cgccgccatc
     2701 gtcgggggca ccgtggcagg catcgtgctg atcggcattc tcctgctggt catctggaag
     2761 gctctgatcc acctgagcga cctccgggag tacaggcgct ttgagaagga gaagctcaag
     2821 tcccagtgga acaatgataa tccccttttc aagagcgcca ccacgacggt catgaacccc
     2881 aagtttgctg agagtggcgg accggtcgcc accatggtga gcaagggcga ggagctgttc
     2941 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     3001 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     3061 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     3121 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3181 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3241 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3301 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3361 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3421 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3481 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     3541 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3601 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     3661 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3721 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3781 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3841 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3901 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3961 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4021 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4081 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4141 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4201 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4261 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4321 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4381 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4441 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4501 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4561 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4621 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4681 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4741 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4801 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4861 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4921 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4981 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5041 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5101 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5161 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5221 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5281 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5341 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5401 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5461 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5521 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5581 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5641 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5701 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5761 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5821 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5881 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5941 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6001 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6061 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6121 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6181 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6241 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6301 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6361 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6421 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6481 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6541 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6601 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6661 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6721 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6781 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6841 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6901 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6961 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag ccc
//
