LOCUS       sequence-249879-        4530 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 4530)
  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 4530)
  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_32_neg_Control
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        346..491
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          569..701
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     684..703
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      755..764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(761..763,764..766,767..1477)
                     /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(806..827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1067..1086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1414..1435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1654..1775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1665..1684)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1719..1738
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2023..2040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2194..2782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2274..2293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2953..3744,3745..3813))
                     /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     3576..3595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3814..3918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3945..4400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4032..4051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4242..4263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttgc
      181 caggagctgg atcgatgata tccgatcgat cgtagcacct gttggctagc ccgggctcga
      241 gatctgcgat ctaagtaagc ttgctgcttc atccccgtgg cccgttgctc gcgtttgctg
      301 gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg
      361 cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc
      421 aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc
      481 gacccgctta acagcgtcaa cagcgtgccg cagatcccaa gcttgcggcc gcttaagctg
      541 cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
      601 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
      661 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact cccaggttca
      721 atacagctct taagcggccg caagctagct tgccgccacc atggtgagca agggcgagga
      781 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      841 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      901 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
      961 cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1021 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1081 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1141 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1201 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1261 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1321 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     1381 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1441 cgccgggatc actctcggca tggacgagct gtacaagtga gcctctagac tccctcagcg
     1501 gcgcgccgct ctccagaaac ttcgcaaaca cctatgccta acgccagccg aatcctgcag
     1561 tccgttacct agtcgaaagg cattctggga gcatcgcgtc acagatctca cgtgtctaga
     1621 gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     1681 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1741 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1801 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1861 gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc
     1921 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     1981 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
     2041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     2401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     2641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4381 aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     4441 atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat
     4501 taaggtacgg gaggtacttg gagcggccgc
//
