LOCUS       sequence-227831-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG:gfp:P2A:mycNRNX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  TITLE     SYNPLA plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7154)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: jk95
FEATURES             Location/Qualifiers
     source          1..7154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        701..978
                     /label=chicken beta-actin promoter
     primer_bind     1258..1280
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1342..1361
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1394..1403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1400..1402,1403..1405,1406..2116)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1400..1402,1403..1405,1406..2116)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1445..1466)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1706..1725)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2053..2074
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2126..2182
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2327..2356
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3548..4136
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3601..3621)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4019..4030)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4179..4300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4190..4209)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4244..4263
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4462..4591
                     /label=AAV2 ITR
     primer_bind     complement(4610..4627)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4610..4626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4619..4641)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4768..5223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4855..4874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5065..5086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5249..5353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5354..5422,5423..6214)
                     /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(5572..5591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6385..6973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6874..6893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatcgta ccattgacgt caataatgac gtatgttccc
      481 atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt acggtaaact
      541 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      601 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      661 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      721 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      781 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      841 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      901 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      961 cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc
     1021 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     1081 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     1141 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     1201 cggctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1261 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1321 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1381 aagaattgga tctgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1441 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1501 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1561 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1621 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1681 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1741 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1801 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1861 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1921 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1981 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2041 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2101 ggacgagctg tacaagggaa gcggagctac taacttcagc ctgctgaagc aggctggaga
     2161 cgtggaggag aaccctggac ctatgtacca gaggatgctc cggtgcggcg ccgatctggg
     2221 atcgcccggg ggcggcagtg gcggcggcgc aggggggcgc ctggccctga tctggatagt
     2281 cccgctcacc ctcggcggcc tcctaggagt ggcctggggg gaattcgaac aaaaactcat
     2341 ctcagaagag gatctagcat ccagtttggg agcgcaccac atccaccatt tccatggcag
     2401 cagcaagcat cattcagtgc ctattgcaat ctacaggtca ccagcatcct tgcgaggcgg
     2461 acacgctggg acgacgtata tctttagcaa aggtggtgga caaattacat ataagtggcc
     2521 tcctaatgac cgacccagta cacgagcaga caggctggcc atcgggttta gcactgttca
     2581 gaaggaagca gtgttggtgc gtgtggacag ttcctcaggc ctgggtgact accttgagct
     2641 gcacatacac caaggaaaaa ttggagttaa atttaatgtt gggacagatg acatcgccat
     2701 tgaggagtcc aatgcaatca ttaatgatgg aaaataccat gtagtgcgtt tcacgaggag
     2761 tggtggcaat gccacgttac aggtggacag ctggccagtc atcgaacgat accctgcagg
     2821 gcgtcagctc acaatcttca atagccaagc aaccataata attggcggga aagagcaggg
     2881 ccagcccttc cagggccagc tctctgggct ttactacaat ggcttgaaag ttctgaatat
     2941 ggcggcagag aacgatgcca acatcgccat agtgggaaat gtgaggctgg tcggtgaagt
     3001 gccttcctct atgacaactg agtcgacagc cactgccatg cagtccgaga tgtccacctc
     3061 aatcatggag accaccacaa ccctggctac cagcacagct cgacgaggaa agccccccac
     3121 aaaggagcct atcagccaga ccacggatga tatccttgtg gcctcggcag agtgtcccag
     3181 cgacgatgag gacattgacc cctgtgagcc gagctcaggt gggttagcca accccacccg
     3241 agtgggcggc cgcgaaccat acccaggctc ggcagaggtg atccgggagt ctagcagtac
     3301 cactggcatg gtggtgggga ttgtcgcagc agcagctctg tgcatcctca tcctcctcta
     3361 tgccatgtac aagtacagga accgggatga agggtcgtac cacgtggatg agagtcgaaa
     3421 ctacatcagt aactcagcac agtccaatgg ggctgtggtc aaggagatgc aacccagcag
     3481 tgctaaaagc gccaacaaaa acaagaagaa caaggataag gagtattatg tctagaagct
     3541 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3601 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3661 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3721 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3781 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3841 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3901 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3961 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4021 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4081 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4141 ataccgtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4201 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4261 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4321 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     4381 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     4441 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     4501 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4561 gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg
     4621 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4681 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4741 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     4801 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4861 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4921 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4981 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5041 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5101 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5161 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5221 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5281 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5341 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5401 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5461 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5521 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5581 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5641 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5701 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5761 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5821 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5881 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5941 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6001 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6061 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7141 cgaggaagcg gaag
//
