LOCUS       Exported                5614 bp ds-DNA     circular SYN 16-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    CMV::SypHy A4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  AUTHORS   Granseth B, Odermatt B, Royle SJ, Lagnado L
  TITLE     Clathrin-mediated endocytosis is the dominant mechanism of vesicle 
            retrieval at hippocampal synapses.
  JOURNAL   Neuron. 2006 Sep 21. 51(6):773-86.
  PUBMED    16982422
REFERENCE   2  (bases 1 to 5614)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 16, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5614
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      602..611
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1166..1876
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1193..1221)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1816..1836
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2402..2523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2439..2458)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2493..2512
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2530..2985)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2667..2688)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2879..2898
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3012..3116
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3118..3475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3141..3161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3326..3461
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3388..3407
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3510..4304
                     /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(3564..3583)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4174..4193
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4492..4511)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4536..4583
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4912..5500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5401..5420
                     /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 gctagcctcg
      601 agccaccatg gacgtggtga atcagctggt ggctgggggt cagttccggg tggtcaagga
      661 gccccttggc ttcgtgaagg tgctgcagtg ggtctttgcc atcttcgcct ttgctacgtg
      721 tggcagctac accggggagc ttcggctgag cgtggagtgt gccaacaaga cggagagtgc
      781 cctcaacatc gaagttgaat tcgagtaccc cttcaggctg caccaagtgt actttgatgc
      841 accctcctgc gtcaaagggg gcactaccaa gatcttcctg gttggggact actcctcgtc
      901 ggctgaattc tttgtcaccg tggctgtgtt tgccttcctc tactccatgg gggccctggc
      961 cacctacatc ttcctgcaga acaagtaccg agagaacaac aaagggccta tgatggactt
     1021 tctggctaca gccgtgttcg ctttcatgtg gctagttagt tcatcagcct gggccaaagg
     1081 cctgtccgat gtgaagatgg ccacggaccc agagaacatt atcaaggaga tgcccatgtg
     1141 ccgccagaca gggaacaccg gtggaagtaa aggagaagaa cttttcactg gagttgtccc
     1201 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
     1261 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
     1321 acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcaat gcttttcaag
     1381 atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
     1441 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa
     1501 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
     1561 tggaaacatt cttggacaca aattggaata caactataac gatcaccagg tgtacatcat
     1621 ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca acattgaaga
     1681 tggaggcgtt caactagcag accattatca acaaaatact ccaattggcg atgggcccgt
     1741 ccttttacca gacaaccatt acctgtttac aacttctact ctttcgaaag atcccaacga
     1801 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
     1861 ggatgaacta tacaaaaccg gtcgccagac agggaacaca tgcaaggaac tgagggaccc
     1921 tgtgacttca ggactcaaca ccacagtggt gtttggcttc ctgaacctgg tgctctgggt
     1981 tggcaactta tggttcgtgt tcaaggagac aggctgggca gccccattca tgcgcgcacc
     2041 tccaggcgcc ccggaaaagc aaccagcacc tggcgatgcc tacggcgatg cgggctacgg
     2101 gcagggcccc ggaggctatg ggccccagga ctcctacggg cctcagggtg gttatcaacc
     2161 cgattacggg cagccagcca gcggtggcgg tggctacggg cctcagggcg actatgggca
     2221 gcaaggctat ggccaacagg gtgcgcccac ctccttctcc aatcagatgt aatctgcaga
     2281 ttaatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2341 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2401 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2521 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2581 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2641 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2701 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2761 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2821 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2881 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2941 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3121 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3481 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3541 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3601 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3661 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3721 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3781 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3841 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3901 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3961 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4021 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4081 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4141 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4201 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4261 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4321 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4441 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4501 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4561 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4621 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4681 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4741 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4801 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4861 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4921 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4981 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5041 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5101 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5161 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5221 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5281 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5341 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5401 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5461 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5521 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5581 tgattctgtg gataaccgta ttaccgccat gcat
//
