LOCUS       Exported                6440 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pOPTN-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  AUTHORS   Park BC, Shen X, Samaraweera M, Yue BY
  TITLE     Studies of optineurin, a glaucoma gene: Golgi fragmentation and cell
            death from overexpression of wild-type and mutant optineurin in two 
            ocular cell types.
  JOURNAL   Am J Pathol. 2006 Dec . 169(6):1976-89.
  PUBMED    17148662
REFERENCE   2  (bases 1 to 6440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6440
                     /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"
     CDS             963..974
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2386..3105
                     /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             2386..3102
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2431..2452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2692..2711)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3039..3060
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3228..3349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3265..3284)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3319..3338
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3356..3811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3493..3514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3705..3724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3838..3942
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3944..4301
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3967..3987)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4152..4287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4214..4233
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4336..5130
                     /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(4390..4409)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5000..5019
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5318..5337)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5362..5409
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5738..6326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6227..6246
                     /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 attcccacca tggctatgtc ccatcaacct
      661 ctcagctgcc tcactgaaaa ggaggacagc cccagtgaaa gcacaggaaa tggacccccc
      721 cacctggccc acccaaacct ggacacgttt accccggagg agctgctgca gcagatgaaa
      781 gagctcctga ccgagaacca ccagctgaaa gaagccatga agctaaataa tcaagccatg
      841 aaagggagat ttgaggagct ttcggcctgg acagagaaac agaaggaaga acgccagttt
      901 tttgagatac agagcaaaga agcaaaagag cgtctaatgg ccttgagtca tgagaatgag
      961 aaattgaagg aagagcttgg aaaactaaaa gggaaatcag aaaggtcatc tgaggacccc
     1021 actgatgact ccaggcttcc cagggccgaa gcggagcagg aaaaggacca gctcaggacc
     1081 caggtggtga ggctacaagc agagaaggca gacctgttgg gcatcgtgtc tgaactgcag
     1141 ctcaagctga actccagcgg ctcctcagaa gattcctttg ttgaaattag gatggctgaa
     1201 ggagaagcag aagggtcagt aaaagaaatc aagcatagtc ctgggcccac gagaacagtc
     1261 tccactggca cggcattgtc taaatatagg agcagatctg cagatggggc caagaattac
     1321 ttcgaacatg aggagttaac tgtgagccag ctcctgctgt gcctaaggga agggaatcag
     1381 aaggtggaga gacttgaagt tgcactcaag gaggccaaag aaagagtttc agattttgaa
     1441 aagaaaacaa gtaatcgttc tgagattgaa acccagacag aggggagcac agagaaagag
     1501 aatgatgaag agaaaggccc ggagactgtt ggaagcgaag tggaagcact gaacctccag
     1561 gtgacatctc tgtttaagga gcttcaagag gctcatacaa aactcagcga agctgagcta
     1621 atgaagaaga gacttcaaga aaagtgtcag gcccttgaaa ggaaaaattc tgcaattcca
     1681 tcagagttga atgaaaagca agagcttgtt tatactaaca aaaagttaga gctacaagtg
     1741 gaaagcatgc tatcagaaat caaaatggaa caggctaaaa cagaggatga aaagtccaaa
     1801 ttaactgtgc tacagatgac acacaacaag cttcttcaag aacataataa tgcattgaaa
     1861 acaattgagg aactaacaag aaaagagtca gaaaaagtgg acagggcagt gctgaaggaa
     1921 ctgagtgaaa aactggaact ggcagagaag gctctggctt ccaaacagct gcaaatggat
     1981 gaaatgaagc aaaccattgc caagcaggaa gaggacctgg aaaccatgac catcctcagg
     2041 gctcagatgg aagtttactg ttctgatttt catgctgaaa gagcagcgag agagaaaatt
     2101 catgaggaaa aggagcaact ggcattgcag ctggcagttc tgctgaaaga gaatgatgct
     2161 ttcgaagacg gaggcaggca gtccttgatg gagatgcaga gtcgtcatgg ggcgagaaca
     2221 agtgactctg accagcaggc ttaccttgtt caaagaggag ctgaggacag ggactggcgg
     2281 caacagcgga atattccgat tcattcctgc cccaagtgtg gagaggttct gcctgacata
     2341 gacacgttac agattcacgt gatggattgc atcattcggg atccaatggt gagcaagggc
     2401 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2461 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2521 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2581 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2641 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2701 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2761 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2821 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2881 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2941 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3001 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3061 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     3121 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3181 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3241 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3301 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3361 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3421 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3481 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3541 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3601 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3661 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3721 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3781 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3841 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3901 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3961 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4021 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4081 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4141 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4201 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4261 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4321 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4381 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4441 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4501 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4561 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4621 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4681 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4741 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4801 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4861 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4921 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4981 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5041 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5101 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5161 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5221 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5281 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5341 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5401 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5461 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5521 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5581 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5641 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5701 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5761 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5821 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5881 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5941 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6001 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6061 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6121 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6181 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6241 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6301 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6361 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6421 accgtattac cgccatgcat
//
