LOCUS       sequence-152836-        7483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  AUTHORS   Cetin A, Callaway EM
  TITLE     Optical control of retrogradely infected neurons using 
            drug-regulated ""TLoop"" lentiviral vectors.
  JOURNAL   J Neurophysiol. 2014 May;111(10):2150-9. doi: 10.1152/jn.00495.2013.
            Epub 2014 Feb 26.
  PUBMED    24572099
REFERENCE   2  (bases 1 to 7483)
  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"; doi: 
            "10.1152/jn.00495.2013"; journalName: "J Neurophysiol"; date: 
            "2014-05"; volume: "111"; issue: "10"; pages: "2150-9"
COMMENT     Sequence Label: pEIAV-iSyn-eGFPW
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1877..2324
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          2438..2913
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      2993..3002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2999..3001,3002..3004,3005..3718)
                     /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(2999..3001,3002..3004,3005..3715)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3044..3065)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3305..3324)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3652..3673
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3920..4508
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3973..3993)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4391..4402)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      4905..5360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4992..5011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5202..5223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5429..5448)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5548..5570
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5608..5626)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5694..5798
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5799..5867,5868..6659)
                     /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(6017..6036)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6830..7418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7319..7338
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
       61 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
      121 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
      181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
      421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
      481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      541 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      661 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgggcactc agattctgcg gtctgagtcc cttctctgct
      781 gggctgaaaa ggcctttgta ataaatataa ttctctactc agtccctgtc tctagtttgt
      841 ctgttcgaga tcctacagtt ggcgcccgaa cagggacctg agaggggcgc agaccctacc
      901 tgttgaacct ggctgatcgt aggatccccg ggacagcaga ggagaactta cagaagtctt
      961 ctggaggtgt tcctggccag aacacaggag gacaggtaag tagggagacc ctttgacatg
     1021 gagcaaggcg ctcaagaagt tagagaaggt gacggtacaa gggtctcaga aattaactac
     1081 tggtaactgt aattgggcgc taagtctagt agacttattt catgatacca actttgtaaa
     1141 agaaaaggac tggcagctga gggatgtcat tccattgctg gaagatgtaa ctcagacgct
     1201 ggaattcggg tattgtagaa agggcaaata ggaccttgaa agagaagatt caaagtcata
     1261 gagacaacac tcaaacactg gaggcagctt tacaacttgc tctcattact tgtaacaaag
     1321 ggagggaaag tatgggagga cagacaccat gggaagtatt tatcactaat caagcacaag
     1381 taatacatga gaaactttta ctacagcaag cacaatcctc caaaaaattt tgtttttaca
     1441 aaatccctgg tgaacatgat tggaagggac ctactagggt gctgtggaag ggtgatggtg
     1501 cagtagtagt taatgatgaa ggaaagggaa taattgctgt accattaacc aggactaagt
     1561 tactaataaa gccaaattga gtattgttgc atgcatttga tatatgggat tatttggtaa
     1621 aggggtaaca tggtcagcat cgcattctat ggggggatcc cagggggaat ctcaacccct
     1681 attacccaac agtcagaaaa atctaagtgt gaggagaaca caatgtttca accttattgt
     1741 tataataatg acagtaagaa cagcatggca gaatcgaagg aagcaagaga ccaagaaatg
     1801 aacctgaaag aagaatctaa agaagaaaaa agaagaaatg actggtggaa aaaagctgca
     1861 gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg
     1921 cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
     1981 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
     2041 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
     2101 tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc
     2161 accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga
     2221 taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt
     2281 ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgagg cgcgccgtga
     2341 aagggcgaat tccagcacac tggcggccgt tactagtgga tccgagctcg gtaccaagct
     2401 ttcacgcgcg ccgagctcgg atcctgagaa cttcagggtg agtctatggg acccttgatg
     2461 ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag
     2521 ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct
     2581 tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc
     2641 agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat
     2701 aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg
     2761 taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct
     2821 tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta
     2881 atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg
     2941 ctggcccatc actttggcaa aggtcgacgg taccgcgggc ccgggatcca tcgccaccat
     3001 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3061 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3121 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3181 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3241 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3301 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3361 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3421 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3481 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3541 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3601 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3661 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     3721 cggccgcgag catcttaccg ccatttattc ccatatttgt tctgtttttc ttgatttggg
     3781 tatacatttg aatgtcaata aaacaaaatg gtggggcaat catctacatt tcatgggata
     3841 tgtgattact agttcaggtg tattgccaca agacaaacat gttaagaaaa tttcccgtta
     3901 tttgcactct gttcctgtta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3961 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     4021 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     4081 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     4141 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     4201 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     4261 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     4321 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4381 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4441 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4501 ctccccgcct gtttcgcctc ggcgtccggt ccgtgttgct tggtcttcac ctgtgcagac
     4561 ttgcgaacca tggattccac cgtgaacttt gtctcctggc atgcaaatcg tcaacttggc
     4621 atgccaaatc gattagaaaa acaagggggg aactgtgggg tttttatgag gggttttata
     4681 caattgggca ctcagattct gcggtctgag tcccttctct gctgggctga aaaggccttt
     4741 gtaataaata taattctcta ctcagtccct gtctctagtt tgtctgttcg agatcctaca
     4801 gagctcatgc cttggttaat taaggagatc cgggctggcg taatagcgaa gaggcccgca
     4861 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     4921 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4981 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5041 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagagctt tacggcacct
     5101 cgaccgcaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5161 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5221 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5281 ttcggcctat tggttaaaaa atgagctgat ttaacaaata tttaacgcga attttaacaa
     5341 aatattaacg tttacaattt cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5401 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5461 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5521 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5581 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5641 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5701 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5761 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5821 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5881 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5941 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6001 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6061 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6121 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6181 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6241 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6301 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6361 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6421 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6481 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6541 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6601 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6661 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6721 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6781 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6841 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6901 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6961 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7021 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7081 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7141 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7201 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7261 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7321 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7381 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7441 tacggttcct ggccttttgc tggccttttg ctcacatggc tcg
//
