LOCUS       sequence-209811-        5767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses porcine annexin A1-GFP fusion protein for live cell 
            imaging, all type II Ca2+ binding sites deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5767)
  AUTHORS   Rescher U, Zobiack N, Gerke V
  TITLE     Intact Ca(2+)-binding sites are required for targeting of annexin 1 
            to endosomal membranes in living HeLa cells.
  JOURNAL   J Cell Sci. 2000 Nov;113 ( Pt 22):3931-8.
  PUBMED    11058080
REFERENCE   2  (bases 1 to 5767)
  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: "J Cell 
            Sci."; date: "2000-11"; volume: "113 ( Pt 22)"; pages: "3931-8"
COMMENT     Sequence Label: cmamxA1-GFP
FEATURES             Location/Qualifiers
     source          1..5767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(696..707)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1647..1656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1653..1655,1656..1658,1659..2372)
                     /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(1653..1655,1656..1658,1659..2369)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1698..1719)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1959..1978)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2306..2327
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2495..2616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2532..2551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2586..2605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2623..3078)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2760..2781)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2972..2991
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3105..3209
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3211..3568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3211..3402
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3234..3254)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3419..3554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3603..4397
                     /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             3603..4397
                     /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(3657..3676)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4267..4286
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4585..4604)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4629..4676
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5005..5593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5494..5513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcaaaaat ggcaatggta tctgagttcc
      601 ttaagcaggc ctggtttatt gacaatgaag agcaagaata cattaaaact gtgaaaggat
      661 ccaaaggtgg tcctgggtca gcagtgagcc cctatcctac cttcaatcca tcctcagatg
      721 ttgaagcctt gcataaagcg ataacagtga aaggtgtgga tgaagcaacc atcattgaaa
      781 ttctaactaa gagaaccaat gcacagcgtc agcagatcaa ggcagcatat ctccaggaga
      841 aaggaaagcc cctggatgaa gctctgaaga aagccctcac aggtcacctt gaggaagttg
      901 ctttggcttt gctgaaaact ccagctcagt ttgatgctga cgaactccgt gctgccatga
      961 agggccttgg aactgatgaa gacactctga atgaaatttt ggcatcaaga actaacagag
     1021 aaatcagaga aattaacaga gtgtataaag aagaactgaa gagagatctg gctaaagaca
     1081 ttacctcagc cacatctgga gattatcaga aggctttgct ttctcttgct aagggtgacc
     1141 gatctgagga ccttgctata aatgatgact tggctgatac agatgcaagg gctttatatg
     1201 aagcaggaga acgaagaaaa gggacagatt tgaatgtgtt cattaccatt cttactacca
     1261 gaagctatcc tcatcttcgc agagtgtttc agaagtattc caagtacagc aagcatgaca
     1321 tgaacaaagt tctggacctg gcgttgaaag gtgacatcga gaattgcctt acagttgttg
     1381 tgaagtgtgc tacaagcaaa ccaatgttct ttgctgagaa gcttcatcag gccatgaagg
     1441 gtattggaac tcgtcataag acactgatca ggattatggt ttcccgttct gaaattgaca
     1501 tgaatgacat caaagcatgc tatcagaagt tgtatggtat ctccctctgc caagccatcc
     1561 tggatgcaac caagggagat tatgaaaaaa ttctggttgc tctatgtgga ggagacgtcg
     1621 acggtaccgc gggcccggga tccatcgcca ccatggtgag caagggcgag gagctgttca
     1681 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1741 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1801 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1861 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1921 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1981 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2041 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2101 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2161 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2221 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2281 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2341 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     2401 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2461 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2581 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2641 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2701 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2761 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2821 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2881 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2941 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3001 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3061 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3121 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3181 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3241 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3301 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3361 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3421 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3481 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3541 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3601 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3661 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3721 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3781 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3841 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3901 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3961 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4021 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4081 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4141 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4201 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4261 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4321 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4381 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4441 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4501 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4561 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4621 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4681 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4741 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4801 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4861 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4921 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4981 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5041 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5101 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5161 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5221 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5281 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5341 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5401 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5461 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5521 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5581 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5641 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5701 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5761 agttccg
//
