LOCUS       sequence-466615-        5650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP cargo with 50bp NOLC homology arms.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  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 5650)
  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: pDY1440 STITCHR GFP NOLC1 payload
FEATURES             Location/Qualifiers
     source          1..5650
                     /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
     intron          1674..2690
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2614..2636
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             2823..3488
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             2823..3488
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     primer_bind     complement(3578..3595)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3584..3808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3880..3897)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3880..3896)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3889..3911)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4038..4466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4125..4144)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4335..4356
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4519..4623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4624..4692,4693..5484)
                     /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(4842..4861)
                     /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 catgttcatg ccttcttctt tttcctacag atccttaata
     2701 ttcaatgaag cgcgggtaaa cggcgggagt aactatgact ctcttaaggt ctagttacaa
     2761 ctggtcctga gtcgtgctgc gtcgacaacg gtagtgacgc gtattgcctg gaggccgcca
     2821 ccatgcccgc catgaagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg
     2881 agctggtggg cggcggagag ggcacccccg agcagggccg catgaccaac aagatgaaga
     2941 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct
     3001 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca
     3061 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga
     3121 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg
     3181 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg
     3241 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc
     3301 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca
     3361 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc
     3421 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg
     3481 ccttcgccag atctcgagct cgagcggacg ccggcattcg ccgcgtggtt cccagcgacc
     3541 tgtatcccct cgtcctaggt cagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     3601 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3661 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3721 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3781 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg
     3841 agatccacta gttctagcct cgaggctaga gcggccgcca ctggccgtcg ttttacaacg
     3901 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3961 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4021 cctgaatggs saayskracg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4081 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4141 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4201 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4261 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4321 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4381 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4441 gatttaacaa aaatttaacg cgaattttaa caaaatatkm wmsmkkrcrm ktymsrtkss
     4501 mcwtttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4561 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4621 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4681 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4741 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4801 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4861 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4921 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4981 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5041 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5101 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5161 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5221 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5281 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5341 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5401 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5461 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5521 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5641 atcaaaggat
//
