LOCUS       sequence-62342-e        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  AUTHORS   Cherry TJ, Wang S, Bormuth I, Schwab M, Olson J, Cepko CL
  TITLE     NeuroD factors regulate cell fate and neurite stratification in the 
            developing retina.
  JOURNAL   J Neurosci. 2011 May 18;31(20):7365-79. doi: 
            10.1523/JNEUROSCI.2555-10.2011.
  PUBMED    21593321
REFERENCE   2  (bases 1 to 7287)
  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: 
            "10.1523/JNEUROSCI.2555-10"; journalName: "J Neurosci"; date: 
            "2011-05-18- 18"; volume: "31"; issue: "20"; pages: "7365-79"
COMMENT     Sequence Label: CAG-NeuroD2iresGFP
FEATURES             Location/Qualifiers
     source          1..7287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     misc_feature    join(2919..3493,3494..3496,3497..3505)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3086..3103)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3313..3332
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3506..3508,3509..3511,3512..4225)
                     /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(3506..3508,3509..3511,3512..4222)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3551..3572)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3812..3831)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4159..4180
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4311..4330)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4376..4431
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4430..4449)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4790..4806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4790..4806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4803..4825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4814..4830
                     /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)."
     promoter        complement(join(4838..4844,4845..4862,4863..4868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4883..4904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4963..5158
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5009..5144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5164..5298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5201..5220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5255..5274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5366..5383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5537..6125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5617..5636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6296..7087,7088..7156))
                     /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     6919..6938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7157..7261)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attctgagcc gccaccatgc
     1741 tgacccgcct gttcagcgag cccggcctcc tctcggacgt gcccaagttc gccagctggg
     1801 gcgacggcga cgacgacgag ccgaggagcg acaagggcga cgcgccgccg cagcctcctc
     1861 ctgctcccgg gtcgggggct ccaggacccg cccgggccgc caagccagtg tctcttcgtg
     1921 gaggagaaga gatccctgaa cccacgttgg ctgaggtcaa ggaggaagga gagctgggcg
     1981 gcgaggagga ggaggaagag gaggaggagg aaggactgga cgaggcggaa ggcgagcggc
     2041 ccaagaagcg cgggccgaag aaacgcaaga tgaccaaggc gcgtctggag cgctccaagc
     2101 tgcggcgaca gaaggccaac gcgcgggagc gcaaccgcat gcacgacctg aacgcggctc
     2161 tggacaacct gcgcaaggtg gtgccctgct actccaagac gcagaagctg tccaagatcg
     2221 agaccctgcg cctggccaag aactacatct gggctctctc ggagatcttg cgctccggga
     2281 agcggccgga tctggtgtcc tacgtgcaga ctctgtgcaa ggggctgtca cagcccacca
     2341 cgaatctggt ggccggctgc ctgcagctaa actctcgtaa cttcctcacg gagcagggcg
     2401 cggacggcgc cggccgcttt cacggctcgg gtggcccgtt cgccatgcat ccgtacccat
     2461 acccgtgctc ccgcctggca ggcgcacagt gtcaggcggc tggcggcctg ggcggaggcg
     2521 cggcgcacgc cctgcggacc cacggctact gcgccgccta cgagacgctg tacgcggcgg
     2581 ccggtggcgg cggcgctagc ccggactaca acagctccga gtacgagggt ccactcagtc
     2641 ccccgctctg tctcaacggc aacttctcgc tcaagcagga ctcgtccccc gatcacgaga
     2701 agagctacca ctactctatg cactactcgg cgctgcccgg ctcacggccc acgggccacg
     2761 ggctggtctt cggctcgtcg gccgtgcgcg ggggcgtcca ctccgagaat ctcttgtctt
     2821 acgatatgca ccttcaccac gatcggggcc ccatgtacga ggagctcaac gcatttttcc
     2881 ataactgagc ggccgctacg taaattccgc cccccccccc cctctccctc ccccccccct
     2941 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     3001 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3061 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3121 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3181 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3241 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3301 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3361 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3421 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3481 acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt
     3541 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3601 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3661 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3721 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3781 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3841 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3901 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3961 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     4021 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4081 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4141 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     4201 cggcatggac gagctgtaca agtaaagcgg ccggccgcca gcacagtggt cgatcgacct
     4261 gcaggcatgc aagcttcagc caatgccctg gctcacaaat accactgaga tctttttccc
     4321 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4381 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4441 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     4501 acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat
     4561 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     4621 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     4681 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     4741 tctcttatga agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     4801 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4861 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4921 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     4981 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5041 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5101 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5161 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5221 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5281 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5701 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     5761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7261 gcacatttcc ccgaaaagtg ccacctg
//
