LOCUS       sequence-257592-        4526 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 4526)
  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 4526)
  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_66_neg_Control
FEATURES             Location/Qualifiers
     source          1..4526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        57..202
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          280..412
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     395..414
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      466..475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(472..474,475..477,478..1191)
                     /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(517..538)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(778..797)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1125..1146
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1362..1483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1373..1392)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1427..1446
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1731..1748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1902..2490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1982..2001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2661..3452,3453..3521))
                     /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     3284..3303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3522..3626)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3653..4108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3740..3759)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3950..3971
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4239..4287
                     /note="synthetic polyadenylation signal"
     misc_feature    4301..4392
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4341..4360
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga
       61 cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc
      121 ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc
      181 gaccctgcag cgacccgctt aacagcgtca acagcgtgcc gcagatccca agcttgcggc
      241 cgcttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga
      301 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
      361 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac
      421 tcccaggttc aatacagctc ttaagcggcc gcaagctagc ttgccgccac catggtgagc
      481 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      541 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      601 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      661 accctgacct ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      721 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      781 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      841 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      901 tacaacgcca tcagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
      961 gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1021 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1081 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1141 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg agcctctaga
     1201 ctccctcagc ggcgcgccac tggtacgaga acggtggagt acctttcgag cgccggtaag
     1261 aagcgtcgcc tgactaaagg ttcgggtgca gaccagttcc gacacgcgta agatctcacg
     1321 tgtctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     1381 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1441 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1501 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1561 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
     1621 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     1681 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     1741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2341 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     3661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3841 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3961 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4021 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4081 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     4141 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     4201 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
     4261 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     4321 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     4381 gaacatttct ctatcgatag gtaccgagct cgagttccat gagctggatc gatgatatcc
     4441 gatcgatcgt agcacaggac cgctagcccg ggctcgagat ctgcgatcta agtaagcttg
     4501 ctgcttcatc cccgtggccc gttgct
//
