LOCUS       sequence-257586-        5042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer cloning nanotag reporter vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5042)
  AUTHORS   Williams RM, Candido-Ferreira I, Repapi E, Gavriouchkina D, 
            Senanayake U, Ling ITC, Telenius J, Taylor S, Hughes J, 
            Sauka-Spengler T
  TITLE     Reconstruction of the Global Neural Crest Gene Regulatory Network In
            Vivo.
  JOURNAL   Dev Cell. 2019 Oct 21;51(2):255-276.e7. doi: 
            10.1016/j.devcel.2019.10.003.
  PUBMED    31639368
REFERENCE   2  (bases 1 to 5042)
  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.devcel.2019.10"; journalName: "Dev Cell"; date: 
            "2019-10-21- 21"; volume: "51"; issue: "2"; pages: "255-276.e7"
COMMENT     Sequence Label: pTK_Citrine_Nanotag_24_positive_Control
FEATURES             Location/Qualifiers
     source          1..5042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        459..604
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          682..814
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     797..816
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(874..876,877..879,880..1590)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(919..940)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1180..1199)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1527..1548
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1763..1884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1774..1793)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1828..1847
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2132..2149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2303..2891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2383..2402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3062..3853,3854..3922))
                     /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     3685..3704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3923..4027)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4054..4509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4141..4160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4351..4372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4640..4688
                     /note="synthetic polyadenylation signal"
     misc_feature    4702..4793
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4742..4761
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ttccctggcc tattctccta agtaattaga ttttaacagg attttcaaca gatcagagca
       61 atctctgtgg gtgccaatag gggactctcc ctctggcctt accagcaggg cagcatggat
      121 aacatccagg ctgggagtta ataggacaat tactggcctc ttttaaatga gttacattaa
      181 tgcacttaag ctaaatggct ctgcatatga ttaaatatta acatctgcaa aatatgaaat
      241 catgatatgc aatcaagttt tgttgtttta atttcccaaa tgccggattt tggtggtgta
      301 tgtaaatgtg cattggatag tgtatgttaa actgttggct agcccgggct cgagatctgc
      361 gatctaagta agcttgctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt
      421 ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg
      481 tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca
      541 cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc
      601 ttaacagcgt caacagcgtg ccgcagatcc caagcttgcg gccgcttaag ctgcagaagt
      661 tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa
      721 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
      781 cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt tcaatacagc
      841 tcttaagcgg ccgcaagcta gcttgccgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     1081 atgtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag tgagcctcta gactccctca gcggcgcgcc
     1621 tgtcgaagct aaaagtgctc gagcccaata ccatttcgga atctcgcgtg gcgttagtat
     1681 gccactcgat ggtaggggaa cgcagaatgc cagatctcac gtgtctagag tcggggcggc
     1741 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1801 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1861 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1921 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     1981 gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     2041 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     2101 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2161 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2221 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2281 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2701 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3001 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3061 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3121 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3181 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3241 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3301 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3361 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3421 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3481 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3541 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3601 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3661 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3721 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3781 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3841 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3901 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3961 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4021 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     4081 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4141 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4201 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4261 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4321 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4381 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4441 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4501 ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4561 cctcttcgct attacgccag cccaagctac catgataagt aagtaatatt aaggtacggg
     4621 aggtacttgg agcggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     4681 tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa
     4741 actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tctatcgata
     4801 ggtaccgagc tcgagttgcc aggagctttc caccaacact taacctttat gaagaaccaa
     4861 attattttgt tccatgacta agaaaaatct ggcgatatat aattaaaaca acaaagaaga
     4921 gtcccattct atacaagtgt aattacgttc atgtggtatc actgtaattg aaacacagaa
     4981 gatgtttcag tctgtattaa aagagaattt ctttcagtgg gtttcctgaa gctcattaga
     5041 ta
//
