LOCUS       sequence-257583-        5045 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 5045)
  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 5045)
  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_Cerulean_Nanotag_91_positive_Control
FEATURES             Location/Qualifiers
     source          1..5045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          78..210
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     193..212
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      264..273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(270..272,273..275,276..989)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(315..336)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(576..595)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     923..944
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1162..1283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1173..1192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1227..1246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1531..1548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1702..2290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1782..1801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2461..3252,3253..3321))
                     /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     3084..3103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3322..3426)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3453..3908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3540..3559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3750..3771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4039..4087
                     /note="synthetic polyadenylation signal"
     misc_feature    4101..4192
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4141..4160
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4900..5045
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
ORIGIN
        1 cagcgtcaac agcgtgccgc agatcccaag cttgcggccg cttaagctgc agaagttggt
       61 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
      121 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
      181 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccaggttcaa tacagctctt
      241 aagcggccgc aagctagctt gccgccacca tggtgagcaa gggcgaggag ctgttcaccg
      301 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      361 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      421 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt
      481 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      541 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      601 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      661 tcaaggagga cggcaacatc ctggggcaca agctggagta caacgccatc agcgacaacg
      721 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
      781 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      841 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag
      901 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      961 ctctcggcat ggacgagctg tacaagtgag cctctagact ccctcagcgg cgcgccggat
     1021 cctcataaga ccagtggtgc ctaggcaggc tctacggttc cagtactacc caaccatacg
     1081 ctgacgtttg tgcccgttgt aacgcacctg agatctcacg tgtctagagc cggggcggcc
     1141 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1201 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1261 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1321 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     1381 ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     1441 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     1501 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2341 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2401 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2461 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2521 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2581 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2641 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2701 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2761 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2821 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2881 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2941 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3001 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3061 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3121 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3181 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3241 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3301 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3421 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     3481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3661 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3721 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3781 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3841 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3901 tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3961 ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga
     4021 ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     4081 ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa
     4141 ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgatag
     4201 gtaccgagct cgagttccat ggagctttcc accaacactt aacctttatg aagaaccaaa
     4261 ttattttgtt ccatgactaa gaaaaatctg gcgatatata attaaaacaa caaagaagag
     4321 tcccattcta tacaagtgta attacgttca tgtggtatca ctgtaattga aacacagaag
     4381 atgtttcagt ctgtattaaa agagaatttc tttcagtggg tttcctgaag ctcattagat
     4441 attccctggc ctattctcct aagtaattag attttaacag gattttcaac agatcagagc
     4501 aatctctgtg ggtgccaata ggagactctc cctctggcct taccagcagg gcagcatgga
     4561 taacatccag gctgggagtt aataggacaa ttactggcct cttttaaatg agttacatta
     4621 atgcacttaa gctaaatggc tctgcatatg attaaatatt aacatctgca aaatatgaaa
     4681 tcatgatatg caatcaagtt ttgttgtttt aatttcccaa atgccggatt ttggtggtgt
     4741 atgtaaatgt gcattggata gtgtatgtta aaaggaccgc tagcccgggc tcgagatctg
     4801 cgatctaagt aagcttgctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg
     4861 tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc
     4921 gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc
     4981 acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg
     5041 cttaa
//
