LOCUS       sequence-354652-        6031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of GFP under the CAMK2A promoter in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6031)
  AUTHORS   Nehme R, Zuccaro E, Ghosh SD, Li C, Sherwood JL, Pietilainen O, 
            Barrett LE, Limone F, Worringer KA, Kommineni S, Zang Y, 
            Cacchiarelli D, Meissner A, Adolfsson R, Haggarty S, Madison J, 
            Muller M, Arlotta P, Fu Z, Feng G, Eggan K
  TITLE     Combining NGN2 Programming with Developmental Patterning Generates 
            Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission.
  JOURNAL   Cell Rep. 2018 May 22;23(8):2509-2523. doi: 
            10.1016/j.celrep.2018.04.066.
  PUBMED    29791859
REFERENCE   2  (bases 1 to 6031)
  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: 
            "10.1016/j.celrep.2018.04"; journalName: "Cell Rep"; date: 
            "2018-05-22- 22"; volume: "23"; issue: "8"; pages: "2509-2523"
COMMENT     Sequence Label: DRH-307
FEATURES             Location/Qualifiers
     source          1..6031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(5..989)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      1060..1515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1147..1166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1357..1378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1541..1645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1646..1714,1715..2506)
                     /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(1864..1883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2677..3265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3166..3185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3419..3436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3553..3574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3589..3594,3595..3612,3613..3619)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3627..3643
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3632..3654
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3651..3667
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3651..3667
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3686..3706
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3688..3706
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3819..3953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3856..3875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3910..3929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3991..5940)
                     /codon_start=1
                     /product="transposase of the Tol2 transposon from the 
                     medaka fish Oryzias latipes (Kawakami, 2007)"
                     /label=Tol2 transposase
                     /translation="MEEVCDSSAAASSTVQNQPQDQEHPWPYLREFFSLSGVNKDSFKM
                     KCVLCLPLNKEISAFKSSPSNLRKHIERMHPNYLKNYSKLTAQKRKIGTSTHASSSKQL
                     KVDSVFPVKHVSPVTVNKAILRYIIQGLHPFSTVDLPSFKELISTLQPGISVITRPTLR
                     SKIAEAALIMKQKVTAAMSEVEWIATTTDCWTARRKSFIGVTAHWINPGSLERHSAALA
                     CKRLMGSHTFEVLASAMNDIHSEYEIRDKVVCTTTDSGSNFMKAFRVFGVENNDIETEA
                     RRCESDDTDSEGCGEGSDGVEFQDASRVLDQDDGFEFQLPKHQKCACHLLNLVSSVDAQ
                     KALSNEHYKKLYRSVFGKCQALWNKSSRSALAAEAVESESRLQLLRPNQTRWNSTFMAV
                     DRILQICKEAGEGALRNICTSLEVPMFNPAEMLFLTEWANTMRPVAKVLDILQAETNTQ
                     LGWLLPSVHQLSLKLQRLHHSLRYCDPLVDALQQGIQTRFKHMFEDPEIIAAAILLPKF
                     RTSWTNDETIIKRGMDYIRVHLEPLDHKKELANSSSDDEDFFASLKPTTHEASKELDGY
                     LACVSDTRESLLTFPAICSLSIKTNTPLPASAACERLFSTAGLLFSPKRARLDTNNFEN
                     QLLLKLNLRFYNFE"
     regulatory      5937..5946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5983..6001)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5984..6001)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ggcggttccc taaacgagca ttgcttatat agacctccca ttaggcacgc ctaccgccca
       61 tttacgtcaa tggaacgccc atttgcgtca ttgcccctcc ccattgacgt caatggggat
      121 gtacttggca gccatcgcgg gccatttacc gccattgacg tcaatgggag tactgccaat
      181 gtaccctggc gtacttccaa tagtaatgta cttgccaagt tactattaat agatattgat
      241 gtactgccaa gtgggccatt taccgtcatt gacgtcaata gggggcgtga gaacggatat
      301 gaatgggcaa tgagccatcc cattgacgtc aatggtgggt ggtcctattg acgtcaatgg
      361 gcattgagcc aggcgggcca tttaccgtaa ttgacgtcaa tgggggaggc gccatatacg
      421 tcaataggac cgcccatatg acgtcaatag gaaagaccat atatagagac cattgacgtc
      481 aatgggggag tggctatggg cggtattagg aagccccata tatggtatat gggaccgccc
      541 attgggaggg gctatctacg tcaataggaa aacccatata tggaatacta tatggcatag
      601 ggccaataca tagtattgaa cctggccaat agccatattg gcatagggcc atattggata
      661 ttgcctatat attgatcctg gcatatagcc aatatggccg ccattattgg caccatgcca
      721 attcaatatg gcggacttgg cacagggcca agacccctcc tcaaacttgg caccgtgcca
      781 agtagacccc tccccagttg gcacagtgcc aggtccgcca tattgaattg gcatggtgcc
      841 aagtccgcca tattgaattg gctggggtcc agatccacca tattgaattg gcacgagtcc
      901 atatacgcca tattgaattg gcacaggtcc agatccacca tattgaattg gcatgagtcc
      961 atatacgcca tattgaattg gcggtcgact ggcgtaatag cgaagaggcc cgcaccgatc
     1021 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     1081 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1141 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1201 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1261 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1321 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1381 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1441 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1501 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1621 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1681 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1741 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1801 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1861 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1921 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1981 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2041 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2101 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2161 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2221 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2281 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2341 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2401 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2461 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2821 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3301 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3361 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3421 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3481 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3541 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     3601 gctcccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     3661 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctggg
     3721 taccgggccc aatgcattgg cgccgcggcc gcgaattaaa aaacctccca cacctccccc
     3781 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3841 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3901 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctacgtaa
     3961 tacgactcac tatagttcta gaggcgcgcc ctactcaaag ttgtaaaacc tcagatttaa
     4021 cttcagtaga agctgattct caaaattgtt agtgtcaagc ctagctcttt tggggctgaa
     4081 aagcaatcct gcagtgctga aaagcctctc acaggcagcc gatgcgggaa gaggtgtatt
     4141 agtcttgata gagaggctgc aaatagcagg aaacgtgagc agagactccc tggtgtctga
     4201 aacacaggcc agatatccat ccaactcttt gctggcttca tgtgttgtcg gtttcaaaga
     4261 agcgaaaaaa tcttcatcat cagatgaact gttggccaat tccttcttgt ggtccaaagg
     4321 ctccagatgc actctgatgt agtccatgcc tcgttttatg atggtttcat catttgtcca
     4381 agaggtccga aatttaggga gaaggatggc agctgctatg atctcaggat cttcaaacat
     4441 atgcttgaat cgtgtttgga ttccttgttg tagggcatcc acaagtgggt cacagtacct
     4501 gagagaatgg tggagtcgct gaagtttcaa gcttaactga tggacactag gcagcagcca
     4561 ccccagctgt gtattcgttt ccgcttgcaa gatgtcgagt acttttgcaa ctggacgcat
     4621 tgtgttggcc cactctgtca agaacagcat ttctgctgga ttaaacattg gaacctcaag
     4681 agaggtgcat atattccgaa gtgcgccttc tcctgcttct ttgcaaattt gaagaattct
     4741 gtcaacagcc ataaaagttg aattccaccg cgtttggttt ggccttaaaa gctgaagccg
     4801 gctttctgat tcaacagctt cagctgctag agccgatcgg ctgcttttat tccataaagc
     4861 ttggcatttg ccaaagacag atctgtagag tttcttgtag tgttcatttg agagagcttt
     4921 ttgggcatca acgcttgaga ctaggttaag taagtgacag gcacactttt gatgttttgg
     4981 tagctggaat tcgaagccat cgtcttggtc caggactcgt gaggcatctt ggaattccac
     5041 accatcactt ccctcaccac agccttcaga atcagtgtca tcactttcac accttcttgc
     5101 ctcagtctcg atatcattgt tttccacacc aaaaactctg aaagccttca taaagttgga
     5161 accactgtct gtggttgtgc aaacaacctt gtcacgtatt tcatactctg agtggatatc
     5221 attcatggca ctggccagta cctcaaaagt atgagagccc attaatcttt tgcaggcaag
     5281 tgcagcggaa tgtctttcaa gacttccagg gttgatccag tgagcagtta caccaatgaa
     5341 tgactttcta cgtgcagtcc aacaatccgt tgtggttgca atccattcaa cttcactcat
     5401 ggcagcagtc actttctgtt tcatgatcag agcagcttca gctatcttgg agcgtaaagt
     5461 aggccttgta atgacagaaa tgccaggctg cagtgtacta atcagctctt taaatgatgg
     5521 cagatcaaca gtgctgaaag gatgaagtcc ttgaatgatg taccttaata tagctttgtt
     5581 cacagtgact ggagacacat gtttgactgg gaaaactgag tcaactttca gttgcttact
     5641 gctggaagca tgggtggagg tcccgatctt tctcttctgt gctgtcaatt tagagtagtt
     5701 tttgaggtaa tttgggtgca ttctctcaat atgcttcctt aggtttgatg gcgaactttt
     5761 gaaggccgat atttctttat taagcgggag acagaggaca catttcatct tgaatgaatc
     5821 tttatttaca ccacttaaag aaaagaattc gcgaagatac ggccacgggt gctcttgatc
     5881 ctgtggctga ttttggactg tgctgctcgc agctgctgat gaatcacata cttcctccat
     5941 ggtggccggc cggatcctgc aaaaagaaca agtagcttgt attctatagt gtcacctaaa
     6001 tcaagcttgc tccgacgtcc ccaggcagaa t
//
