LOCUS       sequence-275704-        4263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription for mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4263)
  AUTHORS   Borsos M, Perricone SM, Schauer T, Pontabry J, de Luca KL, de Vries 
            SS, Ruiz-Morales ER, Torres-Padilla ME, Kind J
  TITLE     Genome-lamina interactions are established de novo in the early 
            mouse embryo.
  JOURNAL   Nature. 2019 May;569(7758):729-733. doi: 10.1038/s41586-019-1233-0. 
            Epub 2019 May 22.
  PUBMED    31118510
REFERENCE   2  (bases 1 to 4263)
  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: 
            "10.1038/s41586-019-1233-0"; journalName: "Nature"; date: "2019-05";
            volume: "569"; issue: "7758"; pages: "729-733"
COMMENT     Sequence Label: pRN3P_EGFP_m6A_Tracer
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           45..87
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      95..104
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(101..103,104..106,107..817)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(101..103,104..106,107..817)
                     /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(146..167)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(407..426)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     754..775
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1197..1213)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1197..1213)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     3'UTR           1239..1364
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1452..1471)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1453..1471)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1478..1495)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1478..1494)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1487..1509)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1636..2091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1773..1794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1985..2004
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2118..2222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2223..2291,2292..3083)
                     /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(2441..2460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3254..3842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3743..3762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3996..4013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4130..4151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4166..4171,4172..4189,4190..4196)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4204..4220
                     /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     4209..4231
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4228..4244
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4228..4244
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaattaaccc tcactaaagg gaacaaaagc tggagctaag cttgcttgtt ctttttgcag
       61 aagctcagaa taaacgctca actttggcag atctgccgcc atggtgagca agggcgagga
      121 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      181 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      241 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      301 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      361 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      421 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      481 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      541 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
      601 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
      661 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
      721 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
      781 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca
      841 agcttcgaat tctccttcta aaggaaggat atttcttgtg caagatggac aagttagaga
      901 tccagaaaaa gttacaaaag aatttaagca aggtttattt ttaaggaaga gctctctgtc
      961 atcaagaggt tggacaatag aaattctaaa ttgtatagat aagatagagg gttcagaatt
     1021 tacccttgaa gatatgtatc gttttgaaag tgacctaaaa aatatctttg ttaagaacaa
     1081 tcatatcaaa gaaaagatta ggcaacagct tcaaatatta agagacaaag aaataataga
     1141 atttaaaggt agaggaaagt atcggaaatt atgagtttaa acccgctgat cagccggatc
     1201 cactagttct agagcggccg cggatctggt taccactaaa ccagcctcaa gaacacccga
     1261 atggagtctc taagctacat aataccaact tacactttac aaaatgttgt cccccaaaat
     1321 gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattcta aaaaaaaaaa
     1381 aaaaaaaaaa aaaaaaaaaa aamccccccc cccccccccc tgcagggcca agtcggccgg
     1441 tacccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     1501 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1561 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1621 tgaatggcga atggcaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1681 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     1741 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     1801 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     1861 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     1921 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1981 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2041 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2101 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2221 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2341 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2401 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2461 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2521 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2641 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2761 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2821 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3061 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4021 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4081 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4141 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4201 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     4261 tcg
//
