LOCUS       sequence-443181-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-13_Ami transposase
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  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 6401)
  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-hAT-13_AMi
FEATURES             Location/Qualifiers
     source          1..6401
                     /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(2589..2607)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2590..2607)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2627..2644)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2633..2857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2903..3331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2990..3009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3200..3221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3340..3360)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3345..3674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3525..3660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3587..3606
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3840..3961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3877..3896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3931..3950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4010..4026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4010..4026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4023..4045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4034..4050
                     /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(4058..4064,4065..4082,4083..4088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4241..4258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4412..5000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4492..4511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5171..5962,5963..6031))
                     /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     5794..5813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6032..6136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6211..6230)
                     /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 ggagaacgaa aacgtggagt taccgggcct gggtgaagaa gaagcgggca gcagcttaag
      781 ccgtagctta agcccggata tgcgctcaac catgagcagc tgttcacgcc agagcagctt
      841 tctgagcagc tttgtggata aaatgagcaa agaggatcag gagaaggcgg atgaaacctt
      901 agcgcgcgcg atttttgcaa gcggcacccc tttaagcatc accgagaacc agtactggaa
      961 ggaacactat aagctgattc gccctagcta caaactgccg agccgttacc gtttaagcca
     1021 tagcctgctg gatcgcgaat atgaccgcgt gcagaccatg gttatccagc gcatcggtca
     1081 agcagaaagc ctgaccttaa tgagcgatgg ctggaccgat atacaaggca acccgctgct
     1141 gaacattatg ctggcgaccc cggagcctat attcctgaaa gcgattgcga ccaaaagcag
     1201 ccgtcatacc ggcgagtata tcgcgaaact gctgagcgaa gagattgaaa gcgccggccc
     1261 ttcacgtgtt caagcactgg tgaccgacaa tgccagcaac atgaaggccg cctggcagat
     1321 tctgaaggcg aaatacccgc acctgattgt gtttggctgc ctggcccatg gtttaaatct
     1381 gctggcgaag gatattgtga gcgtgaacac catgaaaacc attctggcga actgcaaggc
     1441 cattgtgaaa gtgtttaaca accatcacgt ggccaaccag accctgaaga agctgcagaa
     1501 agaaaagcag ggcaaagaaa gcgctctgct gctgcctggt gaaactcgct ggggtagcgc
     1561 aaccaaatgt ctgcatagcc tgctgcatac caaaaactgc ttacagtgcg cgagcattga
     1621 cgataccgtg agccagatca ttcctgtgaa catccgcaaa cagattctgg acaacgacgt
     1681 gttttgggtg cgtgttcagg gcgcgtttaa cctgctgtta ccggtgagcg tggcgatcaa
     1741 aaaactggaa ggcgataccc cgagcttaag caacatccct gagatttttc gccagctgaa
     1801 cgagaacctg attgaactgc tgcctagcag cccgctgagc aagaaagaag tggtggtgaa
     1861 gcagatgttt accaaacgca gcgagttttg cagccatcct gtgcatctgg cggccaactt
     1921 actggatcct cagcatcgcg gtcaacacct gagcgaagat gaactgagca ccgccctgaa
     1981 caccattatg gaagtggcga agaacgtgcc gaacattgac gaaattgcgg tgatgaccga
     2041 catagcggag tatcgcgcga aagagaaact gtggagcaag gatattattt ggcgcgcggc
     2101 ggagaatatg agccctctga cctggtggaa aggctactgc aacacccgtc tgctgagccg
     2161 tattgcgctg cgcattctga gcattcctcc gactgcggcg tcatgtgaac gtaactggaa
     2221 agcgttcagc agcatcaaaa ccaaaaaacg caaccgcctg accaatgagt gtaccgcgaa
     2281 gctggtgagc attagccaga acctgctgct gctggagaac caaagcgaag cgcagctggt
     2341 gaaaaaaatg caatgcgatg acaagctgcc gctgcctatt gacagcctgg tgaagcctag
     2401 ctttagcgaa cgcgaatgcg gtagcagcag cagcgaatca gatctgccgc agagcgatag
     2461 cgagatggaa ctgattagca gcgccagcga tagcaccgac agcgatagcg aaactggtgg
     2521 tgcagcgatg accagcgtgg aagaaagcta atgagcggcc gctcgagcat gcatctagag
     2581 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     2641 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2701 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2761 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2821 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2881 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3301 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3481 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3541 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3601 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3661 taggcttttg caaaaagctc cccgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3721 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3781 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3961 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6181 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6241 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     6301 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     6361 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
