LOCUS       sequence-444769-        5591 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of Tc1-13_Xt transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 5591)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pCMV-Tc1-13_Xt
FEATURES             Location/Qualifiers
     source          1..5591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1779..1797)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1780..1797)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1817..1834)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1823..2047
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2093..2521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2180..2199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2390..2411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2530..2550)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2535..2864
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2715..2850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2777..2796
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3030..3151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3067..3086)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3121..3140
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3200..3216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3200..3216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3213..3235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3224..3240
                     /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(3248..3254,3255..3272,3273..3278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3293..3314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3431..3448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3602..4190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3682..3701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4361..5152,5153..5221))
                     /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     4984..5003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5222..5326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5401..5420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccgccaccat
      721 ggtgcgcagc aaagaactga gcgaagcctt tcgcaagaag attgtggcgg cgtataagag
      781 cggcaaaggc ttcaaaaaaa tcagcaagga gttcgaaatc agccatagca ccgtgcgcaa
      841 gatcgtgtat aaatggcgca cctttaaaac caccgcgaac atgcctcgtt caggccgccc
      901 tagcaaattt actcctcgcg cggaccgtaa aatgctgaag gaggtgagca agaacccgaa
      961 aatgagcagc cgcgatctgc aacaagcgct ggcgaccgtg gatgttaaag tgcacgcgag
     1021 caccattcgc aaacgcctgc ataactttaa cctgcatggt cgctgtgcgc gccgtaaacc
     1081 gttactgagc aagcgcaaca ttaaagcgcg cctgaagttt gcacgcgaaa acgtggataa
     1141 acaccaggac ttttggaaca acgtgctgtg gaccgacgaa agcaaaatcg aactgttcgg
     1201 ccaccaaaac cgcggtcatg tttggcgtaa accgaacacc gcctttcagg aaaaaaacct
     1261 gattccgacc gtgaaacatg gtggcggtag cgttatggtg tggggttgtt ttgcggcggc
     1321 aggtcctggt cagttaacca tcattgagag caccatgaac agcaccgtgt atcagcgcgt
     1381 gctggaggaa catgtgcgtc cgagcgttcg caaactgaag ctgaaacgca actggaccct
     1441 gcaacacgac aacgacccta agcataccag caaaagcacc aaggattggc tgaagaccaa
     1501 gaagtggcgc gttctggaat ggcctagcca aagccctgat ctgaacccta ttgaaatgct
     1561 gtggggcgat ttaaaacgtg cggtgcacgc acgtaatccg agcaacatta gccagctgaa
     1621 ggagttctgc atcgaagaat ggggcaaact gagcagcgat cgctgtcagc gtttagtgga
     1681 cggctacaaa aaacgcctga ccgcggtgat ttcagcaaaa ggcggcaaca ccagctatta
     1741 atgagcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     1801 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1861 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1921 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1981 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2041 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2221 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2461 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2521 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2581 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2701 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2761 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     2821 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgaaatga
     2881 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     2941 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3001 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     3061 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3121 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     3181 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3241 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3301 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3361 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3421 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3481 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3541 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3601 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3661 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3721 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3781 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3841 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3901 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3961 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4021 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4081 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4141 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4201 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4261 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4381 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4441 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4501 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4561 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4621 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4681 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4741 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4801 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4861 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4921 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4981 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5041 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5101 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5161 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5221 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5281 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5341 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     5401 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     5461 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     5521 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     5581 atatacgcgt t
//
