LOCUS       sequence-443176-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of OposCharlie2 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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"; 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-OposCharlie2
FEATURES             Location/Qualifiers
     source          1..6506
                     /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(2694..2712)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2695..2712)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2732..2749)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2738..2962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3008..3436
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3095..3114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3305..3326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3445..3465)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3450..3779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3630..3765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3692..3711
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3945..4066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3982..4001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4036..4055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4115..4131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4115..4131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4128..4150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4139..4155
                     /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(4163..4169,4170..4187,4188..4193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4208..4229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4346..4363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4517..5105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4597..4616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5276..6067,6068..6136))
                     /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     5899..5918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6137..6241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6316..6335)
                     /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 ggagaacccc aagaacaaaa aaacccgcct gagcaaaggt tcaggtagca gcagcagccg
      781 tccttttcag gaaacgtgga ccgaaatgta tggcatcatc gaaaaaaacg gccgcagctt
      841 ctgtgttctg tgcaccgaaa ccgttgttag ccgcacctgg aacatcaacc gccatttcga
      901 aaccaaccat agccagctgc tgaagaaaag cgaagatgag cgcaaagaat acattagccg
      961 ccagctgcac ctgtataaga gccagagcaa cagcattctg aaatttgtga agggcagcac
     1021 caatttaacc agcgccagct taagcattgc ccatagcatt gcgcagcacg gcaaagcatt
     1081 aagcgaaggc gagttcatta aagaaaccct gctgcgctgt gcacctgttc tgttccacga
     1141 catgcagaac aaagatgcga ttattaagcg catcagcgaa ttacctctga gccgcaacat
     1201 tatcaaggac cagatcatgc gcctgaacac caacgtgcag catcagctga aacgcgatat
     1261 tagcaactgc aaatacttta gcatcagcct ggatgaaacc accgatgtga ccagccatgc
     1321 gcagctggcg attattggcc gctatagcga tggcctgacc atgcgcgaag aactgattaa
     1381 actggtgagc gtgcctacta gcactagcgg cagcgaaatt tgcaaggtgg tgattcagac
     1441 ctttcgcgat ctgagcattg atatcagcaa agtggtgagc gtgaccactg atggtgcgcc
     1501 taacatggtg ggcaaaaaag tgggcttcgt gaaactgttt accgaagcga tcggccatcc
     1561 gatagtgccg tttcattgca ttattcatca ggaagcgctg tgtgcgaaag cgggctttac
     1621 cgatctgaac gatctgatga gcgtggtgac caaaattgtg aacctgattg cggcacgccc
     1681 tctgcataaa cgcgaattta gcgcgctgct gctggaagtg gatagcacct atagcggcct
     1741 gctgatgtac aacaacgtgc gctggctgag ccgtggtaaa gtgctggaac gctttgtgga
     1801 atgctttgag gagatcaaag tgttcctgga agacaaagac ctgggcaact ttcctcagct
     1861 gaacgatgat aaatgggtga acaccctgat gttcttcacc gatctgagcg tgcacattaa
     1921 cgagctgaac ctgaagctgc agggctttgg caagagcatt gatgtgatgt tcggctacat
     1981 caaagcgttt gaaagcaagc tgaagatctt taagcgcgat gtggaaacca aaacctacaa
     2041 atatttccct cgcgtgacca aatactttga aaaagcgagc gccgcggtgc aaaacgagat
     2101 ggaacctctg catatcaaat accagcatgt gctggatagc ctgctggatc agtttagcga
     2161 tcgctttaac cagtttcgca gcctggaaca gaccatgaag atcatcaagt atcctgacgt
     2221 ggtggtgtat agcagcctgg agctgaacgg ctttcagtgg atgcagatcg acgatctgga
     2281 aatgcagctg gcggagttcc aggatagcat ttgggcgcag gtgtttgttg atctgcgcag
     2341 caaactggag aacttagagc gctgccgcct ggaaaaccag gaagaatgcc attatgagca
     2401 ggaaatttgg agcgcgtgga accgtctgcc ggataccttt agcaccctga aaaacattgc
     2461 gatggcgctg ctgaccattt tccctagcac ctacttctgc gaaaccctgt ttagcgcgct
     2521 gaacaacatc aaaaccaaca agcgcaaccg cctgaccgat gaagttagca gcgcgtgttt
     2581 aggcctgaag tgcaccaaat atcagcctag catcgaagat ctggccaacg aaatccagca
     2641 gcagaaaagc cattgatgag cggccgctcg agcatgcatc tagagggccc tattctatag
     2701 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2761 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2821 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2881 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2941 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3001 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3061 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3121 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3181 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3241 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3301 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3361 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3421 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3601 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3661 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3721 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3781 agctccccga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3841 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3901 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     3961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4081 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4141 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6241 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     6301 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     6361 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     6421 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     6481 cgatgtacgg gccagatata cgcgtt
//
