LOCUS       sequence-404684-        8387 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 8387)
  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 8387)
  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 alpha L - mCFP
FEATURES             Location/Qualifiers
     source          1..8387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     284..304
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(329..433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      460..915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(547..566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     757..778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    922..1043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(933..952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     987..1006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1166..1879,1880..1882,1883..1885))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1211..1232)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1560..1579
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1819..1840
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1882..1891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5290..5301)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(5650..5853)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        5854..6157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(6332..6920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6412..6431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7249..7296
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     7321..7340
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(7528..8322)
                     /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             complement(7528..8322)
                     /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(7639..7658)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8249..8268
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(8357..8387,1..327))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8371..8387,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
       61 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
      121 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
      181 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      241 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      301 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
      361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
      481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      901 taacgcttac aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa
      961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1021 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1081 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     1141 attatgatct agagtcgcgg ccgctttact tgtacagctc gtccatgccg agagtgatcc
     1201 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     1261 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     1321 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     1381 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     1441 ggctgatgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1501 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1561 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1621 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     1681 cgccccaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     1741 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     1801 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1861 acagctcctc gcccttgctc accatggtgg cgaccggtcc agcgtcttgg tcaaccttcg
     1921 cgggttccgg agctgcagtt gggagctggc ggctgctctc ttcgtccttg ccaccaccac
     1981 tctcagagtc cttctcatgg aggggcttca ggcagccggg atccccagcc tcttgcccag
     2041 atgccagctg ctcagagtct tctgcagaga ttccattcgg aacacctctg ccagcctcca
     2101 tcttctcctt caggttccgt ttgaagaaac caaccttgta cagcactatg aaaatgagca
     2161 gcagcagcag cagccccccg atgccgctca gcacgtagag gtagagcatc tgcttctcat
     2221 acaccacgtc aaccttcatg acaacctggg ccagggaggc gttgctgcca tagaggtgga
     2281 aatgcttgct gctgttgaag gagatggaga gggagctgca gaggctgaac atggaagagg
     2341 cctcgatctc tcccaccagc tccagagtcc cgatcacttg gacgaggatc tcctgcctga
     2401 agacaacagg gcagcggaac agggctccgg ggagacaagg ctcagctgca tccgggagcc
     2461 tctccagatc ctcatagtgg cagggcacgg gaggctccat ctgcacgctc cactggtgtg
     2521 tgatgggccc ctcgctggga ggctgtggca ccccaaccac agcctccagg gtgggtatgt
     2581 tgtggtcgtg gatggaaggc tggatcctca cctggtacat gtgcttgact tggtggatct
     2641 tggggccttt gggggtgaaa ctgacataga gtgtggagtc ttcttggtcc tggatgagga
     2701 tgttgatggg gtacaggatg gggatgatgg tagtggctga gttgtcctcc aggaggtctg
     2761 agtcctcatt gttacaggtc acattggcgt gcaattcaac cgagtccccc caggagctgt
     2821 ttaccagtgt attaaacatc atctgcagag caaccgagtg gcctgctttg aagatgggag
     2881 agctcacatt gcaagataat gccctggaca gaagcctgga ctcttcagga agctcctcgc
     2941 agctcacagg tatctggcta tggggcttca gcatctccac cttgcggaag gagagtcccg
     3001 gggggaagtg caggtccagc tggacccagt aagcatcttc ttccaagtta ctcaggctca
     3061 gctccacaga gaggctggca aaagcagtta gacgcagggc tctggatctt gcaggagaga
     3121 aggacactct caagtttgcc tcacacttct tgtcctcccc acagctcttc tcaaaaggga
     3181 tctcccaggt ttccgagtgc agggagggtc tcaggatggg cggtatgtcc ttgccctgcg
     3241 ccctttggtc cctcggtgtc ccttcctcct cccaaagaga gaaattcagg gaaacattga
     3301 tgggggagat gaggtcttga acacataccg ggaaatgaaa tgagaagtca gtgcatgaca
     3361 tgctggtggt gacagctata ttccttctga gttcatgtct ccctcctggg aacaaccccc
     3421 gtcttctggt ccggtggcca tccagctgca gagtgtaagt gagattggca accaggcggc
     3481 cttggaactg ggggatgaga gacttgatct ggaaacagat tgtgatatta actccttctt
     3541 tcatcttgtt actggttgaa taggagcact ccacttcatg cactgggatc tcagctggag
     3601 agaaggacat cagggtgacc atatccacca cgggccggga gctcagcacg atcatctggc
     3661 tctcagcccc cacagccaca tctgccaagc catccccttc aaggtccttc accccatgga
     3721 tggagcgtcc aaaccactga attcctgaga gcacttgggt cccttctatc cgctgacttg
     3781 gctggggact aagccccccg tgcctcccat tgaagatgta cacagccccc tgctcctcca
     3841 gaggggcccc cacagccacg tctaccagcc catcgccgtt gatgtctgtc agagcagtga
     3901 tggcttctcc aaaccgcccg agtgggtagc cggggtcccc ctgcagctct gagacttctt
     3961 caaaccccaa ctgtcttctc tggtagataa acacccggcc tcctctctgc tccccataga
     4021 acagtggggc accaatcagc agcagctctg tctccccatc ttggtccacg tcgacgccac
     4081 acagctcccc accgaaataa gagccaatct gggtcccatg gattgtctgg acctggctcc
     4141 agtgtcctcc gccctgtggc tcttggaaca gcagcactcg gcccatgtgc tggtatcgag
     4201 gggctcccga ggccagcaac gaagtctttt gccgggaggg cagccaggtc acggtgtaac
     4261 ccaaatagcc tgctctcact tctggtgtca atggttcatt cccaataaat gtgtcatcct
     4321 gcaggtctgc cttcaggtca agaaagcccc cagcccagtc cttggctcct actgccccca
     4381 cgactgcatg gcccctgctg aggtcagcac tgatgccgct ggaggacagc tccatgttga
     4441 aggaagtcag gtcctgtttg cttgtgccct caatgacata gatcttcttc tgcagctcag
     4501 tgaatagatc tttcagcttc tcaaatgtgt ccagaatttt cacaaactcg ctcgcgggtt
     4561 ttgatgcaaa tttgtggagg gtctcctgac tctccttggt ctgaaaatgc tttccaatcc
     4621 cgatgatgta gcggatgatg tctttggccg catcgatgtt gccactgtca gtggcctccc
     4681 catccgtgat gatgataagc actttggtgg catctggccg ggcccccagc tcctcccgga
     4741 acacctctgt cgcgacataa ttgatggcac caaaggtatt ggtcaacagc aacatgtgct
     4801 ttacatgctt cagcagagca tcagggtcct tccatttaac ataatctgag aaatcaaatt
     4861 ctgttttgta gcttgtggaa aactgaacag cagcaaactg gtacgaagtg ttgctgagtt
     4921 tcttcatcac atccttcatg aagtccagaa ttttctgaaa ttcatctggc tgcaagctca
     4981 tcgaaccatc aaacagaaat accaggtcta cgttgccctt gatacattcc tgaaaaccag
     5041 ggcgcccctg cagcatggga ccctgcagat tctggcggaa gaggtaacac aggccactca
     5101 gataggtgtt ctggtcacac gttcgagaca gcccagggtc acaggccaaa atgcttccat
     5161 ctgtggggtc tgttgccaag gtcattccca agtacttgga ggtatagttg gaacctctca
     5221 gggtgactgg caggcagtgt cctgtgcccg actggcactg atagaggctt cctgtgctgt
     5281 tcccctcccc tggagctccc acgatgaccc cgtttccgac ctgcaggacg cggtatccaa
     5341 agtgcctccc ggcgcgcggt ggggagaagc tccgcgcgcc ccgcacgtcc aggttgtagc
     5401 tcgaggccgg cgcgaagaaa aagaacccag acagcagcgc catggccatc acagtgatgc
     5461 aggaatcctt catccttcca gcactcgagg gacctgtggc cccaggggca gcgtcaggag
     5521 gcccgtggga tttgctggca acctagacag ggtgaaagag ggaattcgag ctcgcccggg
     5581 gatcctctag aagcttgagc tcgagatctg agtccggtag cgctagcgga tctgacggtt
     5641 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     5701 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     5761 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     5821 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     5881 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     5941 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     6001 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     6061 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     6121 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     6181 aactaatgac cccgtaattg attactatta ataactaatg catggcggta atacggttat
     6241 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     7081 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     7141 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     7201 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     7261 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     7321 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     7381 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     7441 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     7501 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     7561 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     7621 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     7681 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     7741 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     7801 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     7861 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     7921 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     7981 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     8041 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     8101 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     8161 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     8221 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     8281 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     8341 tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcct
//
