LOCUS       sequence-466612-        5240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gluc exon 2 with 50bp AAVS1 homology arms.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5240)
  AUTHORS   Fell CW, Villiger L, Lim J, Hiraizumi M, Tagliaferri D, Yarnall MTN,
            Lee A, Jiang K, Kayabolen A, Krajeski RN, Schmitt-Ulms C, Ramani H, 
            Yousef SM, Roberts N, Vakulskas CA, Nishimasu H, Abudayyeh OO, 
            Gootenberg JS
  TITLE     Reprogramming site-specific retrotransposon activity to new DNA 
            sites.
  JOURNAL   Nature. 2025 Apr 9. doi: 10.1038/s41586-025-08877-4.
  PUBMED    40205048
REFERENCE   2  (bases 1 to 5240)
  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: "Nature. 
            2025 Apr 9. doi: 10.1038/s41586-025-08877-4."
COMMENT     Sequence Label: pDY1656 STITCHR Gluc AAVS1 payload
FEATURES             Location/Qualifiers
     source          1..5240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     494..513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     747..764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    881..902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(917..922,923..940,941..947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    955..971
                     /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)."
     primer_bind     960..982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     979..995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     979..995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1015..1394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1396..1673
                     /label=chicken beta-actin promoter
     primer_bind     2614..2636
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2911..2930
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(3168..3185)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3174..3398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3470..3487)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3470..3486)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3479..3501)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3628..4056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3715..3734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3925..3946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4109..4213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4214..4282,4283..5074)
                     /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(4432..4451)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
       61 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      841 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      901 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      961 gcggataaca atttcacaca ggaaacagct atgaccatga ggcgcgccgg attcgacatt
     1021 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     1081 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     1141 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     1201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1261 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1321 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1381 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     1441 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
     1501 cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg
     1561 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
     1621 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
     1681 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     1741 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     1801 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     1861 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     1921 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     1981 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     2041 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     2101 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     2161 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     2221 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     2281 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     2341 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     2401 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     2461 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     2521 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     2581 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     2641 gctctagagc ctctgctaac catgttcmtg ccywstkcty tkwmykwcar agtgaagaaa
     2701 ttcaatgaag cgcgggtaaa cggcgggagt aactatgact ctcttaaggt ctagttacaa
     2761 ctggacccca cagtggggcc actagggaca ggattggtga cagaaaagcc ccatggatcc
     2821 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
     2881 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt cgatctgtgt
     2941 gtggactgca caactggctg cctcaaaggg cttgccaacg tgcagtgttc tgacctgctc
     3001 aagaagtggc tgccgcaacg ctgtgcgacc tttgccagca agatccaggg ccaggtggac
     3061 aagatcaagg gggccggtgg tgactaagct agcccttagg cctcctcctt cctagtctcc
     3121 tgatattggg tctaaccccc acccctaggt cagagctcgc tgatcagcct cgactgtgcc
     3181 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3241 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3301 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3361 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3421 ctggggctcg agatccacta gttctagcct cgaggctaga gcggccgcca ctggccgtcg
     3481 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3541 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3601 agttgcgcag cctgaatggs smayskracg cgccctgtag cggcgcatta agcgcggcgg
     3661 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3721 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3781 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3841 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3901 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3961 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4021 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatattr wyrcwkrcrr
     4081 yttcmrttsg ccwttymggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4141 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4201 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4261 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4321 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4381 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4441 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4501 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4561 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4621 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4681 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4741 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4801 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4861 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4921 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4981 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5041 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5101 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5161 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5221 cgtagaaaag atcaaaggat
//
