LOCUS       sequence-258414-        4948 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 4948)
  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 4948)
  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_BsmBI_LacZ_Cerulean_Nanotag_91
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1..7,8..25,26..31))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    46..67
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        235..380
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          458..590
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     573..592
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(650..652,653..655,656..1369)
                     /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(695..716)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(956..975)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1303..1324
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1542..1663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1553..1572)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1607..1626
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1911..1928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2082..2670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2162..2181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2841..3632,3633..3701))
                     /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     3464..3483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3702..3806)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3833..4288
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3920..3939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4130..4151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4366..4905)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLPWN
                     SSSVPIDREMFWHLHLHWGQPILLVCFVSFCFDGERMLVLSIHTKNQHTDVMKIKIFYC
                     GRSKYLPYLNITYLSW"
     polyA_signal    4419..4467
                     /note="synthetic polyadenylation signal"
     misc_feature    4481..4572
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4521..4540
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     4755..4777
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4769..4786
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4770..4786
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4796..4815
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4796..4814
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4862..4882)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4862..4880)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4901..4917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4901..4917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4914..4936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4925..4941
                     /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)."
ORIGIN
        1 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
       61 cacattaatt gcgttgcgct cactgcccgc tttccaccgg tcgtctcagg accgctagcc
      121 cgggctcgag atctgcgatc taagtaagct tgctgcttca tccccgtggc ccgttgctcg
      181 cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca
      241 caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg
      301 cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga
      361 ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcccaag cttgcggccg
      421 cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
      481 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
      541 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
      601 ccaggttcaa tacagctctt aagcggccgc aagctagctt gccgccacca tggtgagcaa
      661 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      721 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      781 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      841 cctgacctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
      901 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      961 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1021 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1081 caacgccatc agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1141 caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1201 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1261 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1321 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgag cctctagact
     1381 ccctcagcgg cgcgccggat cctcataaga ccagtggtgc ctaggcaggc tctacggttc
     1441 cagtactacc caaccatacg ctgacgtttg tgcccgttgt aacgcacctg agatctcacg
     1501 tgtctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     1561 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1621 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1681 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1741 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
     1801 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     1861 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     1921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     3841 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3901 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3961 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4021 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4081 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4141 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4201 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4261 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     4321 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     4381 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
     4441 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     4501 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     4561 gaacatttct ctatcgatag gtaccgagct cgagttccat gggagacggg cgccgctaca
     4621 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4681 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4741 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     4801 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt
     4861 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4921 gaaattgtta tccgctcaca attccaca
//
