LOCUS       sequence-243875-        5825 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express copGFP under EF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  TITLE     Oka Lab plasmids
REFERENCE   2  (bases 1 to 5825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AV0.1-BWS-copGFP
FEATURES             Location/Qualifiers
     source          1..5825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        117..1295
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          348..1286
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1243..1263
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1335..1997
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     misc_feature    2083..2671
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2136..2156)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2554..2565)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2692..2773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2729..2748)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2783..2802
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2826..2966
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2826..2955
                     /label=AAV2 ITR
     rep_origin      3041..3496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3128..3147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3338..3359
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3513..3532)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3632..3654
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3692..3710)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3778..3882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3883..3951,3952..4743)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4101..4120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4914..5502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5403..5422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5564..5693
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agggtctcca tacgcgccct gcagtttccc aagctttgca aagatggata aagttttaaa
       61 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct
      121 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      181 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      241 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      301 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      361 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      421 ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
      481 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
      541 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
      601 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
      661 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
      721 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
      781 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
      841 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
      901 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
      961 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     1021 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1081 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     1141 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     1201 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     1261 ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tggatccacc atggagagcg
     1321 acgagagcgg cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg
     1381 tggagttcga gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca
     1441 agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg
     1501 gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg
     1561 ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc
     1621 tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg
     1681 tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg
     1741 ccaccgtgga gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca
     1801 ccttcagcct gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca
     1861 agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg
     1921 tggaggagct gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga
     1981 cccccattgc cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg
     2041 ccggacccgg ctccaccgga tctcgctaag aattctatcg ataatcaacc tctggattac
     2101 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2161 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2221 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2281 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2341 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2401 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2461 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     2521 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     2581 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     2641 agtcggatct ccctttgggc cgcctccccg catcgataag taagtgacta gaacttgttt
     2701 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2761 tttttttcac tgcattctag ttgtggtttg tccaaactca tctgcacgcg tggcgcgtcg
     2821 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2881 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     2941 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     3001 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3061 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     3121 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3181 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3241 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3301 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3361 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     3421 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3481 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3541 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3601 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3661 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3721 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3901 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3961 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4021 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4081 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4141 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4201 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4261 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4321 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4381 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4441 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4501 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4561 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4621 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4681 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4741 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4801 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4861 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4921 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4981 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5041 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5101 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5161 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5221 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5281 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5341 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5401 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5461 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5521 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     5581 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     5641 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     5701 tggagtcgtg acgtgaatta cgtcataggg ttagggaggt cctgtattag aggtcacgtg
     5761 agtgttttgc gacattttgc gacaccatgt ggtcacgctg ggtatttaag cccgagtgag
     5821 cacgc
//
