LOCUS       sequence-258402-        4952 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 4952)
  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 4952)
  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_89
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..25)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(265..284)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     612..633
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    855..976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(866..885)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     920..939
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1224..1241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1395..1983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1475..1494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2154..2945,2946..3014))
                     /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     2777..2796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3015..3119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3146..3601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3233..3252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3443..3464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3679..4218)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLPWN
                     SSSVPIDREMFWHLHLHWGQPILLVCFVSFCFDGERMLVLSIHTKNQHTDVMKIKIFYC
                     GRSKYLPYLNITYLSW"
     polyA_signal    3732..3780
                     /note="synthetic polyadenylation signal"
     misc_feature    3794..3885
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3834..3853
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     4068..4090
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4082..4099
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4083..4099
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4109..4128
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4109..4127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4175..4195)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4175..4193)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4214..4230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4214..4230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4227..4249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4238..4254
                     /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)."
     promoter        complement(join(4262..4268,4269..4286,4287..4292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4307..4328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4496..4641
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4719..4851
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4834..4853
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4905..4914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4911..4913,4914..4916,4917..4952,1..678)
                     /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"
ORIGIN
        1 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
       61 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      121 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc
      181 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      241 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      301 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      361 ggcaacatcc tggggcacaa gctggagtac aacgccatca gcgacaacgt ctatatcacc
      421 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
      481 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      541 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
      601 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      661 gacgagctgt acaagtgagc ctctagactc cctcagcggc gcgcctcgct aagagaactc
      721 cggtacgacg cgatgcgact acagcaggga ccttcaagcg ctcttctcgg gagaatcgct
      781 agtcgcgctt ggataacctt ccgagatctc acgtgtctag agtcggggcg gccggccgct
      841 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      901 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      961 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1021 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     1081 tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     1141 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     1201 tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1501 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1801 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2041 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2281 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3001 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3121 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3181 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3241 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3301 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3361 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3421 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3481 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3541 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3601 tgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     3661 ctattacgcc agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt
     3721 ggagcggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     3781 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
     3841 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
     3901 gctcgagttc catgggagac gggcgccgct acagggcgcg tcccattcgc cattcaggct
     3961 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     4021 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4081 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     4141 cgggcccccc ctcgaggtcc tccagctttt gttcccttta gtgagggtta attgcgcgct
     4201 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4261 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4321 tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtcgtctcag gaccgctagc
     4381 ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc atccccgtgg cccgttgctc
     4441 gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac
     4501 acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg
     4561 gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg
     4621 accctgcagc gacccgctta acagcgtcaa cagcgtgccg cagatcccaa gcttgcggcc
     4681 gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     4741 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     4801 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     4861 cccaggttca atacagctct taagcggccg caagctagct tgccgccacc atggtgagca
     4921 agggcgagga gctgttcacc ggggtggtgc cc
//
