LOCUS       sequence-99464-e        5387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for HA-tagged alpha2-chimaerin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  AUTHORS   Beg AA, Sommer JE, Martin JH, Scheiffele P
  TITLE     alpha2-Chimaerin is an essential EphA4 effector in the assembly of 
            neuronal locomotor circuits.
  JOURNAL   Neuron. 2007 Sep 6;55(5):768-78.
  PUBMED    17785183
REFERENCE   2  (bases 1 to 5387)
  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: "Neuron."; 
            date: "2007-09-6"; volume: "55(5)"; pages: "768-78"
COMMENT     Sequence Label: pNice-HA-alpha2-chimaerin
FEATURES             Location/Qualifiers
     source          1..5387
                     /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"
     CDS             641..667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             641..667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2175..2296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2212..2231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2266..2285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2303..2758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2440..2461)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2652..2671
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2785..2889
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2891..3248
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2891..3082
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2914..2934)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3099..3234
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3283..4077
                     /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             3283..4077
                     /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(3337..3356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3947..3966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4265..4284)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4309..4356
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4685..5273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5174..5193
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcaccatg tacccctacg acgtgcccga
      661 ctacgccgtc gacatggctc tgaccctgtt cgatacagat gaatatagac ctcctgtttg
      721 gaaatcttac ttataccagc tgcagcagga agcccctcac cctcgaaggg tcacctgcac
      781 ttgtgaggta gaaaacagac caaagtatta tggaagagag tatcatggca tgatctctag
      841 agaagagact gaccagctcc tgagtgtggc tgaggggagc tacctcatcc gtgagagcca
      901 gcggcagcca ggaacgtaca ctttggcttt aagatttgga agtcagacca gaaacttcag
      961 gctgtactac gatggaaagc actttgttgg ggagaaacgc tttgagtcca tccacgatct
     1021 ggtgactgat ggcttgatta ctctctatat tgaaaccaag gcagcagaat acattgccaa
     1081 gatgacgata aacccaattt atgagcacat aggatacaca accttaaaca gagagccagc
     1141 atacaaacag cacatggcag tcctgaaaga gacacatgat gagaaagagg ctacaggcca
     1201 ggatggggta tcagagaaaa ggttgactag tcttgttaga agagcaactc tgaaagaaaa
     1261 cgagcaaatt ccaaaatatg aaaagattca caatttcaag gtgcatacat tcagagggcc
     1321 acactggtgt gaatactgtg ccaactttat gtggggtctc attgctcagg gagtgaaatg
     1381 tgcagattgt ggtttgaatg ttcataagca gtgttccaag atggtcccaa atgactgtaa
     1441 gccagacttg aagcatgtca aaaaggtgta cagctgtgac cttacgacgc tcgtgaaagc
     1501 acataccact aagcggccaa tggtggtaga catgtgcatc agggagattg agtctagagg
     1561 tcttaattct gaaggactat accgagtatc aggatttagt gacctaattg aagatgtcaa
     1621 gatggctttc gacagagatg gtgagaaggc agatatttct gtgaacatgt atgaagatat
     1681 caacattatc actggtgcac ttaaactgta cttcagggat ttgccaattc cactcattac
     1741 atatgatgcc taccctaagt ttatagaatc tgccaaaatt atggatccgg atgagcaatt
     1801 ggaaaccctt catgaagcac tgaaactact gccacctgct cactgcgaaa ccctccggta
     1861 cctcatggca catctaaaga gagtgaccct ccacgaaaag gagaatctta tgaatgcaga
     1921 gaaccttgga atcgtctttg gacccaccct tatgagatct ccagaactag acgccatggc
     1981 tgcattgaat gatatacggt atcagagact ggtggtggag ctgcttatca aaaacgaaga
     2041 cattttattt taggcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     2101 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2161 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2221 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2281 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2341 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2401 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2461 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2521 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2581 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2641 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2701 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2761 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2821 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2881 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2941 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3001 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3121 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3181 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3241 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3301 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3361 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3421 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3481 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3541 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3601 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3661 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3721 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3781 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     3841 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3901 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3961 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4021 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4081 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4141 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4201 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4261 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4321 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4381 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4441 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4501 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4561 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4621 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4681 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4741 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4801 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4861 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4921 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4981 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5041 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5101 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5161 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5221 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5281 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5341 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
