LOCUS       sequence-390427-        4306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with IGFBP2 tagged with Clover.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4306)
  AUTHORS   Conway JRW, Dinc DD, Follain G, Paavolainen O, Kaivola J, Bostrom P,
            Hartiala P, Peuhu E, Ivaska J
  TITLE     IGFBP2 secretion by mammary adipocytes limits breast cancer 
            invasion.
  JOURNAL   Sci Adv. 2023 Jul 14;9(28):eadg1840. doi: 10.1126/sciadv.adg1840. 
            Epub 2023 Jul 12.
  PUBMED    37436978
REFERENCE   2  (bases 1 to 4306)
  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: "Sci Adv."; 
            date: "2023-07-14"; volume: "9(28):eadg1840. doi"; pages: " 
            10.1126/sciadv.adg1840. Epub 2023 Jul 12"
COMMENT     Sequence Label: pENTR221-IGFBP2-Clover
FEATURES             Location/Qualifiers
     source          1..4306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     111..128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(287..314)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(406..492)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     541..563
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     555..572
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     556..572
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    588..687
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1718..1720,1721..1723,1724..2434)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1763..1784)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2371..2392
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2450..2549)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2566..2585)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2567..2585)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2590..2606)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2590..2606)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2708..2727)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2719..3528
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2786..2805)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3675..4263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4164..4183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
       61 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
      121 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
      181 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      241 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
      301 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
      361 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
      421 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
      481 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
      541 cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt
      601 tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa
      661 tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga ctggatccgc
      721 caccatgctg ccgagagtgg gctgccccgc gctgccgctg ccgccgccgc cgctgctgcc
      781 gctgctgccg ctgctgctgc tgctactggg cgcgagtggc ggcggcggcg gggcgcgcgc
      841 ggaggtgctg ttccgctgcc cgccctgcac acccgagcgc ctggccgcct gcgggccccc
      901 gccggttgcg ccgcccgccg cggtggccgc agtggccgga ggcgcccgca tgccatgcgc
      961 ggagctcgtc cgggagccgg gctgcggctg ctgctcggtg tgcgcccggc tggagggcga
     1021 ggcgtgcggc gtctacaccc cgcgctgcgg ccaggggctg cgctgctatc cccacccggg
     1081 ctccgagctg cccctgcagg cgctggtcat gggcgagggc acttgtgaga agcgccggga
     1141 cgccgagtat ggcgccagcc cggagcaggt tgcagacaat ggcgatgacc actcagaagg
     1201 aggcctggtg gagaaccacg tggacagcac catgaacatg ttgggcgggg gaggcagtgc
     1261 tggccggaag cccctcaagt cgggtatgaa ggagctggcc gtgttccggg agaaggtcac
     1321 tgagcagcac cggcagatgg gcaagggtgg caagcatcac cttggcctgg aggagcccaa
     1381 gaagctgcga ccaccccctg ccaggactcc ctgccaacag gaactggacc aggtcctgga
     1441 gcggatctcc accatgcgcc ttccggatga gcggggccct ctggagcacc tctactccct
     1501 gcacatcccc aactgtgaca agcatggcct gtacaacctc aaacagtgca agatgtctct
     1561 gaacgggcag cgtggggagt gctggtgtgt gaaccccaac accgggaagc tgatccaggg
     1621 agcccccacc atccgggggg accccgagtg tcatctcttc tacaatgagc agcaggaggc
     1681 tcgcggggtg cacacccagc ggatgcagtg cggccgcatg gtgagcaagg gcgaggagct
     1741 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1801 cagcgtccgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
     1861 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     1921 cgtggcctgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1981 catgcccgaa ggctacgtcc aggagcgcac catctctttc aaggacgacg gtacctacaa
     2041 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2101 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag
     2161 ccacaacgtc tatatcacgg ccgacaagca gaagaacggc atcaaggcta acttcaagat
     2221 ccgccacaac gttgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2281 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagccatc agtccgccct
     2341 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2401 cgggattaca catggcatgg acgagctgta caagtagtaa gatatctaga cccagctttc
     2461 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     2521 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     2581 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     2641 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     2701 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     2761 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2821 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2881 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2941 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     3001 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     3061 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     3121 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     3181 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3241 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     3301 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3361 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3421 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3481 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     3541 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     3601 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     3661 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3721 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3781 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3841 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3901 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3961 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4021 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4081 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4141 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4201 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4261 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgct
//
