LOCUS       sequence-461578-        5610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes a mTFP1-actin fusion construct under the control of a dox 
            inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5610)
  AUTHORS   Pelosse M, Marcia M
  TITLE     biGMamAct: efficient CRISPR/Cas9-mediated docking of large 
            functional DNA cargoes at the ACTB locus
  JOURNAL   Synthetic Biology 2025.02.13
REFERENCE   2  (bases 1 to 5610)
  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: "Synthetic 
            Biology 2025.02.13"
COMMENT     Sequence Label: psLIB TRE mTFP1-actin
FEATURES             Location/Qualifiers
     source          1..5610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      123..711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     612..631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     865..882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1016..1240
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1307..1840)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2029..2034,2035..2051,2052..2057))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2453..2486)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2566..2584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2603..2621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2640..2658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2677..2695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2751..2769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2788..2806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2825..2843
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2865..2889
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3030..3039
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3036..3743
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    5079..5213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5116..5135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5170..5189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5323..5488
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa tggcgaatgg
       61 gacgcgcccg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      241 ttcagcagag cgcagatacc aaatactatt cttctagtgt agccgtagtt aggccaccac
      301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      781 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      841 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
      901 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt
      961 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg
     1021 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     1081 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     1141 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     1201 aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac
     1261 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     1321 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     1381 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     1441 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     1501 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     1561 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     1621 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     1681 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     1741 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     1801 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     1861 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     1921 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     1981 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     2041 gaacaggctt atgtcaactg ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     2101 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     2161 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     2221 ctgtgcacgg atctgccctt gcttcaggag atcggtagac ctcggccgtc gcggcgcttg
     2281 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     2341 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacagct ttgtttgtac
     2401 tatcaacagg ttgaactgct gatcaacaga tcctctacgc ggccgcggta ccataacttc
     2461 gtatagcata cattatacga agttatctgg ttttactagt atcgattcgc gacctactcc
     2521 ggaatattaa tagatcactc gagggatcca agctggccgc acgtctccct atcagtgata
     2581 gagaagtcga cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct
     2641 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     2701 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     2761 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     2821 gagctcccta tcagtgatag agaaggtacc ccctatataa gcagagctcg tttagtgaac
     2881 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     2941 cgatccagcc tccgcggtgg cggccatcac aagtcatacc gtcccaccat cgggcgcgga
     3001 tcccggtccg aagcgcgcgg aattcaaagg ccgccatggt gagcaagggc gaggagacca
     3061 caatgggcgt aatcaagccc gacatgaaga tcaagctgaa gatggagggc aacgtgaatg
     3121 gccacgcctt cgtgatcgag ggcgagggcg agggcaagcc ctacgacggc accaacacca
     3181 tcaacctgga ggtgaaggag ggagcccccc tgcccttctc ctacgacatt ctgaccaccg
     3241 cgttcgccta cggcaacagg gccttcacca agtaccccga cgacatcccc aactacttca
     3301 agcagtcctt ccccgagggc tactcttggg agcgcaccat gaccttcgag gacaagggca
     3361 tcgtgaaggt gaagtccgac atctccatgg aggaggactc cttcatctac gagatacacc
     3421 tcaagggcga gaacttcccc cccaacggcc ccgtgatgca gaagaagacc accggctggg
     3481 acgcctccac cgagaggatg tacgtgcgcg acggcgtgct gaagggcgac gtcaagcaca
     3541 agctgctgct ggagggcggc ggccaccacc gcgttgactt caagaccatc tacagggcca
     3601 agaaggcggt gaagctgccc gactatcact ttgtggacca ccgcatcgag atcctgaacc
     3661 acgacaagga ctacaacaag gtgaccgttt acgagagcgc cgtggcccgc aactccaccg
     3721 acggcatgga cgagctgtac aagggatcag gatcaggatc aatggatgat gatatcgccg
     3781 cgctcgtcgt cgacaacggc tccggcatgt gcaaggccgg cttcgcgggc gacgatgccc
     3841 cccgggccgt cttcccctcc atcgtggggc gccccaggca ccagggcgtg atggtgggca
     3901 tgggtcagaa ggattcctat gtgggcgacg aggcccagag caagagaggc atcctcaccc
     3961 tgaagtaccc catcgagcac ggcatcgtca ccaactggga cgacatggag aaaatctggc
     4021 accacacctt ctacaatgag ctgcgtgtgg ctcccgagga gcaccccgtg ctgctgaccg
     4081 aggcccccct gaaccccaag gccaaccgcg agaagatgac ccagatcatg tttgagacct
     4141 tcaacacccc agccatgtac gttgctatcc aggctgtgct atccctgtac gcctctggcc
     4201 gtaccactgg catcgtgatg gactccggtg acggggtcac ccacactgtg cccatctacg
     4261 aggggtatgc cctcccccat gccatcctgc gtctggacct ggctggccgg gacctgactg
     4321 actacctcat gaagatcctc accgagcgcg gctacagctt caccaccacg gccgagcggg
     4381 aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc cctggacttc gagcaagaga
     4441 tggccacggc tgcttccagc tcctccctgg agaagagcta cgagctgcct gacggccagg
     4501 tcatcaccat tggcaatgag cggttccgct gccctgaggc actcttccag ccttccttcc
     4561 tgggcatgga gtcctgtggc atccacgaaa ctaccttcaa ctccatcatg aagtgtgacg
     4621 tggacatccg caaagacctg tacgccaaca cagtgctgtc tggcggcacc accatgtacc
     4681 ctggcattgc cgacaggatg cagaaggaga tcactgccct ggcacccagc acaatgaaga
     4741 tcaagatcat tgctcctcct gagcgcaagt actccgtgtg gatcggcggc tccatcctgg
     4801 cctcgctgtc caccttccag cagatgtgga tcagcaagca ggagtatgac gagtccggcc
     4861 cctccatcgt ccaccgcaaa tgcttctgac ctacgtcgac gagctcacta gtcgcggccg
     4921 ctttcgaatc tagagcctgc agtctcgaca agcttgtcga gaagtactag aggatcataa
     4981 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5041 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     5101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5161 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac
     5221 tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     5281 gccaccgctg agcaataact atcataaccc ctaggagatc cgaaccagat aagtgaaatc
     5341 tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt
     5401 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
     5461 cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa
     5521 tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca
     5581 cagaatgaaa atttttctgt catctcttcg
//
