LOCUS       sequence-441967-        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-12_AMi transposase
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  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 7166)
  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-12_AMi
FEATURES             Location/Qualifiers
     source          1..7166
                     /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      1339..1348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3354..3372)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3355..3372)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3392..3409)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3398..3622
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3668..4096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3755..3774)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3965..3986
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4105..4125)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4110..4439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4290..4425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4352..4371
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4605..4726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4642..4661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4696..4715
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4775..4791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4775..4791)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4788..4810)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4799..4815
                     /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(4823..4829,4830..4847,4848..4853))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4868..4889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5006..5023)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5177..5765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5257..5276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5936..6727,6728..6796))
                     /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     6559..6578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6797..6901)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6976..6995)
                     /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 gaagcgcacc tatcagagcg gtgcgagcaa acgcaaagag actaaggaaa aaaaagcgcg
      781 cgatgaaaaa ggtcgccaga ccctggatca attcggctgg gtggtgaaag gcaacaagga
      841 agtgggtagc ggtaccggtg atgcaggtgg tacctcactg gcaggcgatg aggaatatag
      901 cgatcacgag aacatcacca aagaagcgag cagcgatgtt gcgggcgaat gcgaaaatga
      961 aagcggcctg gtggtgcaag cggagggtga tattgcgagc aacgaaaaga acgaaaccat
     1021 tgacctggtt tcaggcagcg cgcctgaact gcatcaggaa atgtttagca acgatcctgc
     1081 gacctggggc catttagatc gcctgcagcg cgatctgatt gtgagcaaag gcccgcctcc
     1141 taagcctgca agcttcccgt gcgatagcag caagtggagc tttccgacca gcgtgttttg
     1201 caaaaccctg agcaacgatg agagcgtgaa gcgcgactgg ctgatttgga gccataaggc
     1261 ggcaggcctg ttttgttttc cgtgctgcct gtttcgcgac cagaacaaaa accaggcgct
     1321 gtatcatagc cgcttttgcc atggcggcgt gaatgacaat tggcgcaagc tgtacgataa
     1381 agtgaaaagc catgaagcga acagcggcca cttacagcat tacttacagt ggaaagacct
     1441 ggcgaatgcg ttaagcggca agggtgcgat tgacaacgcc ctggaaaaag gcattcatag
     1501 cgaaaaggag aaatggcgca aagtgctgcg tggcctggtt gatgtgaccc tgtttctggc
     1561 cgaacagaac ttaccgttcc gcggtagcca tagcaccctg gaacatgatg attgcggcct
     1621 gtttctgagc accgtgcagt tcatcagcaa gtataaccag gatctgaact gccacctgga
     1681 aaccgttacc gaacaccgta tggcaggcga acgcatgcaa gcgcattaca tgagctggcg
     1741 cagccagaac gaatatctgg aagtgtgcgc gaaaaaggtg ctgaacacca ttctggaaga
     1801 agtgcgcaac gcgcgctatt atggcattat cgtggatggc accccggatg tttcacatac
     1861 cgagcagctg gtgtttatta tgcgctacgt gcatgagaag gacggcaagt gggatgtgca
     1921 ggaacgcttt ctgaaaatgg tggactgcga caaaaaaaag ggcgtggata ttgtggaaca
     1981 gatcaagaag gtgctggcgg aatgcaaagt ggagctgcaa aactgtcgcg gccagggcta
     2041 tgataacgcc agcaacatga gcggcaagta tcagggcgtg aaaacccgca ttcaggacga
     2101 aaatccgcag gcgctgttta gcccttgtag cgcgcatagc ctgaatctgt gcggtaccca
     2161 cgcgatggaa aagagcccgg aagtgaaaac ctattttggc aacgtgcaga aactgtatac
     2221 cgtgtttagc tgcagccctg cgcgttggaa gatactgcaa gaaaccgcgg gcttaagcct
     2281 gcatagcgtg tcaaccaccc gttggagcag ccgtattgac gctgtgcgtc ctctggtgaa
     2341 gaaccctaaa ggcatgctgg atagcctgtt taagattgag gaagaactgg atctgaccgc
     2401 cgaaacccaa gcagatgtga gcggcctgat tgaatggatg cagagctttg aatttatcct
     2461 gctgaccacc gtgtggtata aaattctgca gagcatcaac attcgcagca agctggtgca
     2521 aggcagcaaa ctgaccatgg aagagggcgt gcagcatgtg aaaagcctgc tggaagaaat
     2581 ccagcgcctg aatgatagct ggccgagctt actgaccgaa gcgaaagtga ttgccgaagg
     2641 cctgggcatt accactgagt tcaagcgcac ccatcgtcgt attcgccgca ccaaacattt
     2701 ccataacgaa cctgccaaca ccgagcatac tatcagcagc ccggaagata aatttcgcgt
     2761 ggaggtgttt attaccgcca tggatatgct gctgagcgat ctgcgtagct tagcggaaac
     2821 caccggtaaa ctgagcagcc tgtttagccc tatcctggac cctcctgcag ttgcagagga
     2881 tgaagaaatt gcgaagttcg cggaaagcct gagcactacc tatcctcgcg acgtgaacaa
     2941 agacgaactg caggaggaac tgtgcctgtt ttacaaaatt cgcaaggaca gcaacttatg
     3001 taccagcggc ggtgcaatgg gtaccgcgtt agaactgctg aaccagatct acggcaaagg
     3061 cgtggaatca gtgttcccgc agatttgcat ttgcctgcgc atgctgatcg cgttacctgt
     3121 tagcgcagcg agcggtgaac gtaccttcag caagctgaaa atcatcaaga actatctgcg
     3181 cagcgcgatc ggtgaagatc gtctgaacca cctgattatg ctgagcgtgg agaacgcact
     3241 ggcgaaacag attaactacg acgatgtgat cgatgatttt gcgagcatga aagcgcgcaa
     3301 aaagtacttc acctgatgag cggccgctcg agcatgcatc tagagggccc tattctatag
     3361 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     3421 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3481 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3541 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3601 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3661 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3721 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3781 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3841 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3901 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3961 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4021 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4081 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4141 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4201 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4261 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4321 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4381 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4441 agctccccga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4501 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4561 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4621 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4681 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4741 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4801 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4861 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4921 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4981 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5041 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5101 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5161 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5221 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5281 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5341 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5401 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5461 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5521 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5581 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5641 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5701 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5761 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5821 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5881 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5941 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6001 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6061 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6121 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6181 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6241 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6301 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6361 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6421 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6481 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6541 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6601 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6661 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6721 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6781 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6841 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6901 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     6961 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7021 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     7081 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     7141 cgatgtacgg gccagatata cgcgtt
//
