LOCUS       sequence-251835-        5929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Left arm of TALEN pair 2 for cleavage of apobb.1 STOP locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5929)
  AUTHORS   Thierer JH, Ekker SC, Farber SA
  TITLE     The LipoGlo reporter system for sensitive and specific monitoring of
            atherogenic lipoproteins.
  JOURNAL   Nat Commun. 2019 Jul 31;10(1):3426. doi: 10.1038/s41467-019-11259-w.
  PUBMED    31366908
REFERENCE   2  (bases 1 to 5929)
  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"; journalName: "Nat 
            Commun."; date: "2019-07-31"; volume: "10(1):3426. doi"; pages: " 
            10.1038/s41467-019-11259-w"
COMMENT     Sequence Label: pT3TS-TAL-apobb.1-PR2-LA
FEATURES             Location/Qualifiers
     source          1..5929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        146..164
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           173..215
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             285..305
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             306..332
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             2634..3221
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     3'UTR           3260..3385
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(3484..3503)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3485..3503)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3510..3527)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3510..3526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3519..3541)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3694..4122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3804..3825)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4016..4035
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4149..4253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4254..4322,4323..5114)
                     /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     complement(4472..4491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5285..5873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5774..5793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgaatta accctcacta aagggaagct tgcttgttct
      181 ttttgcagaa gctcagaata aacgctcaac tttggcagat ctaactcgag aaagatattg
      241 tatatatcgt aacaatagga ggttcaacaa tggcttcctc ccctccaaag aaaaagagaa
      301 aggttagttg gaaggacgca agtggttggt ctagagtgga tctacgcacg ctcggctaca
      361 gtcagcagca gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg
      421 aggcactggt gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg
      481 cagcgttagg gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga
      541 cacacgaaga catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct
      601 tgctcacgga tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg
      661 tgaagattgc aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg
      721 cactgacggg tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaacg
      781 gtggcggsma syrassgsts smmmcskkrm msygrcyscg gaccaagtgg tggctatcgc
      841 cagcaacggt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg
      901 ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa
      961 gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac
     1021 cccggaccaa gtggtggcta tcgccagcaa caatggcggc aagcaagcgc tcgaaacggt
     1081 gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc
     1141 tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt
     1201 gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg
     1261 cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg
     1321 cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga
     1381 aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt
     1441 ggtggctatc gccagcaacg gtggcggcaa gcaagcgctc gaaacggtgc agcggctgtt
     1501 gccggtgctg tgccaggacc atggcctcac acccgatcag gtcgtggcta tcgccagcaa
     1561 cggtggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga
     1621 ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc
     1681 gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga
     1741 ccaagtggtg gctatcgcca gccacgatgg cggcaagcaa gcgctcgaaa cggtgcagcg
     1801 gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc
     1861 cagcaacggt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg
     1921 ccaggaccat ggcctgactc cggaccaagt ggtggctatc gccagccacg atggcggcaa
     1981 gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac
     2041 cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt
     2101 gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc
     2161 tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt
     2221 gctgtgccag gaccatggcc tgactccgga ccaagtggtg gctatcgcca gccacgatgg
     2281 cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg
     2341 cctgaccccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga
     2401 aagcattgtg gcccagctga gccggcctga tccggcgttg gccgcgttga ccaacgacca
     2461 cctcsysgcs ttggccngcs tygrcsracg wccwscycgt cgccttggcc tgcctcggcg
     2521 gacgtcctgc catggatgca gtgaaaaagg gattgccgca cgcgccggaa ttgatcagaa
     2581 gagtcaatcg ccgtattggc gaacgcacgt cccatcgcgt tgcctctaga tcccagctag
     2641 tgaaatctga attggaagag aagaaatctg aacttagaca taaattgaaa tatgtgccac
     2701 atgaatatat tgaattgatt gaaatcgcaa gaaattcaac tcaggataga atccttgaaa
     2761 tgaaggtgat ggagttcttt atgaaggttt atggttatcg tggtaaacat ttgggtggat
     2821 caaggaaacc agacggagca atttatactg tcggatctcc tattgattac ggtgtgatcg
     2881 ttgatactaa ggcatattca ggaggttata atcttccaat tggtcaagca gatgaaatgc
     2941 aaagatatgt cgaagagaat caaacaagaa acaagcatat caaccctaat gaatggtgga
     3001 aagtctatcc atcttcagta acagaattta agttcttgtt tgtgagtggt catttcaaag
     3061 gaaactacaa agctcagctt acaagattga atcatatcac taattgtaat ggagctgttc
     3121 ttagtgtaga agagcttttg attggtggag aaatgattaa agctggtaca ttgacacttg
     3181 aggaagtgag aaggaaattt aataacggtg agataaactt ttaataggct agtgactgac
     3241 taggatctgg ttaccactaa accagcctca agaacacccg aatggagtct ctaagctaca
     3301 taataccaac ttacacttta caaaatgttg tcccccaaaa tgtagccatt cgtatctgct
     3361 cctaataaaa agaaagtttc ttcacattct aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3421 ammccccccc cccccccctg caggtcgact ctactagagg atccccgggc gagctccaat
     3481 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     3541 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3601 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3661 gaayskramk ygymmkskwk mrtattttgt taaaattcgc gttaaatttt tgttaaatca
     3721 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3781 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3841 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3901 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3961 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4021 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4081 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcs mcwttymggs
     4141 krmrygtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctca
//
