LOCUS       sequence-240247-        6130 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 6130)
  AUTHORS   Butler MH, David C, Ochoa GC, Freyberg Z, Daniell L, Grabs D, 
            Cremona O, De Camilli P
  TITLE     Amphiphysin II (SH3P9; BIN1), a member of the amphiphysin/Rvs 
            family, is concentrated in the cortical cytomatrix of axon initial 
            segments and nodes of ranvier in brain and around T tubules in 
            skeletal muscle.
  JOURNAL   J Cell Biol. 1997 Jun 16. 137(6):1355-67.
  PUBMED    9182667
REFERENCE   2  (bases 1 to 6130)
  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 
            Biol. 1997 Jun 16. 137(6):1355-67."
COMMENT     Sequence Label: pEGFPC1-muscle Amphiphysin 2
FEATURES             Location/Qualifiers
     source          1..6130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(716..737)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(977..996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1324..1345
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2976..3097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3013..3032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3067..3086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3104..3559)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3241..3262)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3453..3472
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3586..3690
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3692..4049
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3692..3883
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3715..3735)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3900..4035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4084..4878
                     /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             4084..4878
                     /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(4138..4157)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4748..4767
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5066..5085)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5110..5157
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5486..6074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5975..5994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1381 gtacaagtcc ggactcagat ccatggcaga gatgggcagt aaaggggtga cggcgggaaa
     1441 gatcgccagc aacgtgcaga agaagctcac ccgcgcgcag gagaaggttc tccagaagct
     1501 ggggaaggca gatgagacca aggatgagca gtttgagcag tgcgtccaga atttcaacaa
     1561 gcagctgacg gagggcaccc ggctgcagaa ggatctccgg acctacctgg cctccgtcaa
     1621 agccatgcac gaggcttcca agaagctgaa tgagtgtctg caggaggtgt atgagcccga
     1681 ttggcccggc agggatgagg caaacaagat cgcagagaac aacgacctgc tgtggatgga
     1741 ttaccaccag aagctggtgg accaggcgct gctgaccatg gacacgtacc tgggccagtt
     1801 ccccgacatc aagtcacgca ttgccaagcg ggggcgcaag ctggtggact acgacagtgc
     1861 ccggcaccac tacgagtccc ttcaaactgc caaaaagaag gatgaagcca aaattgccaa
     1921 ggccgaggag gagctcatca aagcccagaa ggtgtttgag gagatgaatg tggatctgca
     1981 ggaggagctg ccgtccctgt ggaacagccg cgtaggtttc tacgtcaaca cgttccagag
     2041 catcgcgggc ctggaggaaa acttccacaa ggagatgagc aagctcaacc agaacctcaa
     2101 tgatgtgctg gtcggcctgg agaagcaaca cgggagcaac accttcacgg tcaaggccca
     2161 gcccagaaag aaaagtaaac tgttttcgcg gctgcgcaga aagaagaaca gtgacaacgc
     2221 gcctgcaaaa gggaacaaga gcccttcgcc tccagatggc tcccctgccg ccacccccga
     2281 gatcagagtc aaccacgagc cagagccggc cggcggggcc acgcccgggg ccaccctccc
     2341 caagtcccca tctcagccag cagaggcctc ggaggtggcg ggtgggaccc aacctgcggc
     2401 tggagcccag gagccagggg agacggcggc aagtgaagca gcctccagct ctcttcctgc
     2461 tgtcgtggtg gagaccttcc cagcaactgt gaatggcacc gtggagggcg gcagtggggc
     2521 cgggcgcttg gacctgcccc caggtttcat gttcaaggta caggcccagc acgactacac
     2581 ggccactgac acagacgagc tgcagctcaa ggctggtgat gtggtgctgg tgatcccctt
     2641 ccagaaccct gaagagcagg atgaaggctg gctcatgggc gtgaaggaga gcgactggaa
     2701 ccagcacaag gagctggaga agtgccgtgg cgtcttcccc gagaacttca ctgagagggt
     2761 cccatgacgg cggggcccag gcagcctccg ggcgtgtgaa gaacacctcc tcccggaatt
     2821 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     2881 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2941 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3001 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3061 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     3121 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3181 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3241 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3301 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3361 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3421 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3481 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3541 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3661 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3721 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3841 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3901 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3961 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4021 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4081 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4141 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4201 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4261 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4321 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4381 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4441 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4501 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4561 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4621 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4681 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4741 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4801 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4861 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4921 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4981 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5041 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5101 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5161 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5221 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5281 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5341 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5401 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5461 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5521 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5581 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5641 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5701 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5761 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5821 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5881 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5941 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6001 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6061 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6121 cttttgctca
//
