LOCUS       sequence-393706-        6591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized Pseudoalteromonas TnsC from a CMV 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6591)
  AUTHORS   Lampe GD, King RT, Halpin-Healy TS, Klompe SE, Hogan MI, Vo PLH, 
            Tang S, Chavez A, Sternberg SH
  TITLE     Targeted DNA integration in human cells without double-strand breaks
            using CRISPR-associated transposases.
  JOURNAL   Nat Biotechnol. 2023 Mar 29. doi: 10.1038/s41587-023-01748-1.
  PUBMED    36991112
REFERENCE   2  (bases 1 to 6591)
  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 
            Biotechnol. 2023 Mar 29. doi: 10.1038/s41587-023-01748-1."
COMMENT     Sequence Label: pSL3713 (pTnsC_Pse)
FEATURES             Location/Qualifiers
     source          1..6591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(105..209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(284..303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        475..854
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        855..1058
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1009..1029
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1103..1122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1103..1121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1169..1178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2219..2239
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2282..2302
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2345..2365
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2384..2402)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2385..2402)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2422..2439)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2428..2652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2698..3126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2785..2804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2995..3016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3135..3155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3140..3469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3320..3455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3382..3401
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3536..4330
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3590..3609)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4200..4219
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4504..4625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4541..4560)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4595..4614
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4674..4690)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4674..4690)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4687..4709)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4698..4714
                     /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(4722..4728,4729..4746,4747..4752))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4767..4788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4905..4922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5076..5664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5156..5175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5835..6591,1..35,36..104))
                     /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     6458..6477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
       61 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcat
      121 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
      181 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
      241 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
      301 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      361 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      421 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      481 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      541 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      601 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      661 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      721 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      781 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      841 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      901 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      961 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     1021 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     1081 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
     1141 gagctcggat ccctacttcc aatccaatgc caccatgggc aacgccctga ccgagatcca
     1201 gatcgagaag ctgaggaatt tcagcgactg catcgtgatg cacccacaga tcaagacaat
     1261 cttcaacgac tttgatgagc tgaggctgaa tcgcaagttt cagtccgacc agcagtgtat
     1321 gctgctgatc ggcgataccg gcgtgggcaa gtctcacaca atcaaccact acaagaagcg
     1381 ggtgctggcc acccagaact attccagaaa tacaatgccc gtgctggtgt cccggatcag
     1441 ccggggcaag ggcctggacg ccaccctggt gcagatgctg gccgatctgg agctgttcgg
     1501 cagctcccag atcaagaagc ggggctacaa gaccgatctg acaaagaagc tggtggagtc
     1561 tctgatcaag gcccaggtgg agctgctgat catcaacgag ttccaggagc tgatcgagtt
     1621 taagtctgtg caggagagac agcagatcgc caatggcctg aagtttatca gcgaggaggc
     1681 caaggtgcct atcgtgctgg tgggcatgcc atgggcagca aagatcgcag aggagccaca
     1741 gtgggcctct aggctggtgc ggaagagaaa gctggagtat ttcagcctga agaacgactc
     1801 caagtacttt cgccagtatc tgatgggcct ggccaagaag atgcctttcg atgtgccccc
     1861 taagctggag agcaagaaca ccacaatcgc cctgtttgca gcatgcaggg gagagaatcg
     1921 cgccctgaag cacctgctgc tggaggccct gaagctggcc ctgagctgta acgagtacct
     1981 ggagaataag cacttcatca ccgcctatga caagtttgat ttctttaacg acaaggagaa
     2041 gctgaagtcc aagaatcctt tcaagcagga catcaaggat atcgaaatct acgaagtgat
     2101 caagaacagc agctataacc caaatgccct ggaccccgag gatatgctga cagatcgggt
     2161 gtttgccatc gtgaagggct ccggcaagcg gactgctgat ggcagtgaat ttgagtcccc
     2221 aaagaagaag agaaaggtgg ggagcggcaa aagaaccgcc gacggcagcg aattcgagtc
     2281 acccaagaag aagaggaaag tcggctccgg aaaacggaca gccgacggaa gcgagttcga
     2341 gtcaccgaaa aagaagcgaa aagtttaatc tagagggccc tattctatag tgtcacctaa
     2401 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2581 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2641 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3121 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3481 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3541 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3601 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3661 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3721 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3781 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3841 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3901 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3961 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4021 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4081 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4141 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4201 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4261 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4321 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4381 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4441 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4501 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4621 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4681 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4741 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4801 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4861 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5701 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5761 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5821 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5941 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6421 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc t
//
