LOCUS       sequence-62344-e        7142 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 7142)
  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 7142)
  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-NeuroD6iresGFP
FEATURES             Location/Qualifiers
     source          1..7142
                     /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(2774..3348,3349..3351,3352..3360)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2941..2958)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3168..3187
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3361..3363,3364..3366,3367..4080)
                     /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(3361..3363,3364..3366,3367..4077)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3406..3427)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3667..3686)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4014..4035
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4166..4185)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4231..4286
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4285..4304)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4645..4661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4645..4661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4658..4680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4669..4685
                     /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(4693..4699,4700..4717,4718..4723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4738..4759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4818..5013
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4864..4999
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5019..5153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5056..5075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5110..5129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5221..5238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5392..5980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5472..5491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6151..6942,6943..7011))
                     /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     6774..6793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7012..7116)
                     /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 attcatgtta acactaccgt
     1741 ttgacgagtc tgtcgtaatg cccgaatccc agatgtgcag aaagtttgct agacaatgtg
     1801 aggaccagaa acaaattaag aaaccagaga gctttccaaa acaagttgtc cttcgaggaa
     1861 agagcattaa aagggcccct ggagaagaaa ccgagaaaga agaggaggaa gaagacagag
     1921 aggaagaaga tgagaatggc ttgtccagaa ggagggggct caggaaaaaa aagaccacca
     1981 aactacgact ggaaagggtc aagttcagga gacaggaagc taatgcgcgc gagaggaacc
     2041 ggatgcacgg cctcaatgat gctctggaca atttgcgaaa agtggtcccc tgttactcta
     2101 aaacccaaaa actgtccaaa atagaaactt tacgactggc caaaaattac atctgggcac
     2161 tttctgaaat tctgaggatt ggcaagagac cggatctgct cacgttcgtc caaaacttat
     2221 gcaaaggtct ttcccagcca actacaaact tggtggcagg ctgcttacag ctcaacgcca
     2281 gaagtttcct gatgggtcag ggtggggagg ctgcccacca cacaaggtca ccctactcca
     2341 cattctaccc accctaccac agccctgagc tggccactcc cccagggcat gggactcttg
     2401 ataattccaa gtccatgaaa ccctacaatt actgcagtgc atatgaatcc ttctatgaaa
     2461 gtacctcccc tgagtgtgcc agccctcagt ttgaaggtcc cttaagtcct cccccaatta
     2521 actataatgg gatattttcc ctgaagcaag aagaaacctt ggactatggc aaaaattaca
     2581 attatggcat gcattactgt gcagtgccac ccaggggtcc ccttgggcag ggtgccatgt
     2641 tcaggttgcc caccgacagc cacttccctt acgacttaca tctgcgcagc caatctctca
     2701 ctatgcaaga tgaattaaat gcagtttttc ataattaatg agggcggccg ctacgtaaat
     2761 tccgcccccc ccccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     2821 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2881 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2941 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     3001 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     3061 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     3121 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     3181 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     3241 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     3301 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     3361 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3421 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3481 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3541 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3601 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3661 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3721 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3781 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3841 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3901 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3961 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4021 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     4081 agcggccggc cgccagcaca gtggtcgatc gacctgcagg catgcaagct tcagccaatg
     4141 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     4201 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     4261 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     4321 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgccatatgc tggctgccat
     4381 gaacaaaggt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     4441 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     4501 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     4561 ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc cctcgacctg
     4621 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4681 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4741 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4801 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4861 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4921 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4981 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     5041 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5101 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     5161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5581 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5821 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6061 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6121 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6181 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6241 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6301 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6361 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6421 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6481 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6541 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6601 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6661 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6721 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6781 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6841 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6901 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6961 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7141 tg
//
