LOCUS       sequence-7330-ex        6557 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 6557)
  AUTHORS   Ting JT, Kelley BG, Sullivan JM
  TITLE     Synaptotagmin IV does not alter excitatory fast synaptic 
            transmission or fusion pore kinetics in mammalian CNS neurons.
  JOURNAL   J Neurosci. 2006 Jan 11. 26(2):372-80.
  PUBMED    16407532
REFERENCE   2  (bases 1 to 6557)
  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 Neurosci.
            2006 Jan 11. 26(2):372-80."
COMMENT     Sequence Label: rSytIV pIE
FEATURES             Location/Qualifiers
     source          1..6557
                     /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"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(1916..2490,2491..2493,2494..2502)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2083..2100)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2310..2329
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2503..2505,2506..2508,2509..3222)
                     /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(2503..2505,2506..2508,2509..3219)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2548..2569)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2809..2828)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3156..3177
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3345..3466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3382..3401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3436..3455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3473..3928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3610..3631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3822..3841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3955..4059
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4061..4418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4061..4252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4084..4104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4269..4404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4453..5247
                     /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             4453..5247
                     /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(4507..4526)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5117..5136
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5435..5454)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5479..5526
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5855..6443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6344..6363
                     /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 gctagcgcgc
      601 caccatggct cctatcacca ccagccgcgt ggaattcgat gaaattccca cagtggtggg
      661 catcttcagt gcttttggcc tggtcttcac tgtgtctctc tttgcctgga tctgctgcca
      721 gagaagatcc gctaagtcca acaagactcc tccatacaag tttgtgcacg tgcttaaagg
      781 agttgatatc tacccagaaa acctaagtag caaaaagaag tttggaggag atgacaagag
      841 tgaagcaaag cgtaaagcag ctttgcctaa cctttccctg caccttgatc tcgagaagcg
      901 agacctcaat ggcaatttcc ccaaaaccaa ccccaaagct ggcagctctt ctgatctgga
      961 aaatgtcacc ccaaagctct ttccggagac ggaaaaggag gccgtctccc ctgagagctt
     1021 gaagtccagc acttccctca cttcagagga gaaacaagag aagctgggca ctctcttctt
     1081 gtctctagag tacaacttcg agaagaaagc atttgtggtg aacatcaagg aagcccaggg
     1141 cttaccagcc atggatgaac aatccatgac ctctgaccca tacatcaaga tgacaatctt
     1201 accagagaag aagcacaaag tgaaaaccag agtgctgagg aagacgctgg accccgtgtt
     1261 tgacgaaacc ttcacattct atggggtccc ttatccccac atccaagagc tgtccctgca
     1321 cttcaccgtc ctgagttttg acaggttctc aagagatgat gtcattggag aagtcctcgt
     1381 tcctctttca gggattgaat tgtcagacgg gaaaatgtta atgaccagag agatcatcaa
     1441 gagaaatgct aagaagtctt ctggacgggg tgaacttctg gtctccctct gttatcagtc
     1501 cactacaaac acgctcactg tggttgtctt aaaagcgcgg cacctaccga aatctgatgt
     1561 gtctggactt tcagatccct acgtcaaagt gaacctgtac catgccaaga agagaatctc
     1621 taaaaagaag actcacgtga aaaagtgcac tcccaacgca gtgttcaacg aactgtttgt
     1681 ctttgatatt ccttgtgaga gtcttgaaga aataagtgtt gaatttttag ttttggattc
     1741 tgaaagggga tcccgaaatg aggtgatcgg gcggttggtc ctgggtgcca cagcagaagg
     1801 aagtggtggg gggcactgga aggagatctg tgacttcccc aggagacaaa ttgctaagtg
     1861 gcatatgctc tgtgatggtt agctgcagtc gacggtaccg cgggcccgat atccgcccct
     1921 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     1981 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2041 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2101 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2161 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2221 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2281 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2341 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     2401 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2461 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     2521 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2581 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2641 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2701 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2761 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2821 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2881 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2941 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3001 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3061 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3121 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3181 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     3241 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3301 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3361 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3421 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3481 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3541 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3601 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3661 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3721 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3781 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3841 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3901 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4021 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4081 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4141 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4261 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4321 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4381 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4441 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4501 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4561 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4621 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4681 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4741 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4801 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4861 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4921 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4981 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5041 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5101 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5161 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5221 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5281 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5341 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5401 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5461 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5521 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5581 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5641 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5701 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5761 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6001 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6481 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6541 gtattaccgc catgcat
//
