LOCUS       sequence-490656-        5943 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  AUTHORS   Skoda RC, Seldin DC, Chiang MK, Peichel CL, Vogt TF, Leder P
  TITLE     Murine c-mpl: a member of the hematopoietic growth factor receptor 
            superfamily that transduces a proliferative signal.
  JOURNAL   EMBO J 1993 Jul;12(7):2645-53.
  PUBMED    8334987
REFERENCE   2  (bases 1 to 5943)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J"; 
            date: "1993-07"; volume: "12(7)"; pages: "2645-53"
COMMENT     Sequence Label: pSK-c-mpl
FEATURES             Location/Qualifiers
     source          1..5943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(132..151)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(133..151)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(158..175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(158..174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(167..189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(316..771)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(453..474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     665..684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        798..902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(903..971,972..1763)
                     /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     complement(1121..1140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1934..2522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2423..2442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2676..2693
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2810..2831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2846..2851,2852..2869,2870..2876)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2884..2900
                     /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)."
     primer_bind     2889..2911
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2908..2924
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2908..2924
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2945..2963
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3000..3016
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3000..3016
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
       61 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaactc gagggggcgg
      121 tacccaatts gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
      181 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      241 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      301 tgaatggcga atggcaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
      361 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      421 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
      481 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
      541 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
      601 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
      661 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
      721 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
      781 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      841 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      901 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      961 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1021 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1081 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1141 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1201 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1261 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1321 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1381 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1441 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1501 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1561 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1621 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1681 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1741 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1801 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1861 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1921 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1981 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2041 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2101 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2161 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2221 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2281 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2341 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2401 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2461 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2521 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2581 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2641 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2701 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2761 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     2821 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     2881 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     2941 tcgaaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct
     3001 ctagaactag tggatccccc gggctgcagg aattcggcac gagcggagaa gatgccctct
     3061 tgggccctct tcatggtcac ctcctgcctc ctcttggccc ttccaaacca ggcacaagtc
     3121 accagccaag atgtcttctt gctggccttg ggcacagagc ccctgaactg cttctcccaa
     3181 acatttgagg acctcacctg cttctgggat gaggaagagg cagcacccag tgggacatac
     3241 cagctgctgt atgcctaccg aggagagaag ccccgtgcat gccccctgta ttcccagagt
     3301 gtgcccacct ttggaacccg gtatgtgtgc cagtttccag cccaggatga agtgcgcctc
     3361 ttctttccgc tgcacctctg ggtgaagaat gtgtccctca accagacttt gatccagcgg
     3421 gtgctgtttg tggatagtgt gggcctgcca gctcccccca gggtcatcaa ggccaggggt
     3481 gggagccaac caggggaact tcagatccac tgggaggccc ctgctcctga aatcagtgac
     3541 ttcctgaggc atgaactccg ctatggcccc acggattcca gcaacgccac tgccccctcc
     3601 gtcattcagc tgctctccac agaaacctgc tgccccactt tgtggatgcc gaacccagtc
     3661 cctgttcttg accagcctcc gtgtgttcat ccgacagcat cccaaccgca tggaccagct
     3721 ccatttctga cagtgaaggg tggaagctgt ctcgtctcag gcctccaggc tagcaaatcc
     3781 tactggctcc agctacgcag ccaacccgac ggggtctccc ttcgtggctc ctggggaccc
     3841 tggtccttcc ctgtgactgt ggatcttcca ggagatgcag tgacaattgg acttcagtgc
     3901 tttaccttgg atctgaagat ggtcacctgc cagtggcagc aacaagaccg cactagctcc
     3961 caaggcttct tccgtcacag caggacgagg tgctgcccca cagacaggga ccccacctgg
     4021 gagaaatgtg aagaggagga accgcgtcca ggatcacagc ccgctctcgt ctcccgctgc
     4081 cacttcaagt cacgaaatga cagtgttatt cacatccttg tagaggtgac cacagcgcaa
     4141 ggtgccgttc acagctacct gggctcccct ttttggatcc accaggctgt gctccttccc
     4201 accccgagcc tgcactggag ggaggtctca agtggaaggc tggagttgga gtggcagcac
     4261 cagtcatctt gggcagctca agagacctgc taccagctcc ggtacacggg agaaggccgt
     4321 gaggactgga aggtgctgga gccatctctc ggtgcccggg gagggaccct agagctgcgc
     4381 ccccgagctc gctacagctt gcagctgcgt gccaggctca acggccccac ctaccaaggt
     4441 ccctggagcg cctggtctcc cccagctagg gtgtccacgg gctccgagac tgcttggatc
     4501 accttggtga ctgctctgct cctggtgctg agcctcagtg cccttctggg cctactgctg
     4561 ctaaagtggc aatttcctgc gcactacagg agactgaggc atgctttgtg gccctcgctt
     4621 ccagacctac accgggtcct aggccagtac ctcagagaca ctgcagccct aagtccttct
     4681 aaggccacgg ttaccgatag ctgtgaagaa gtggaaccca gcctcctgga aatcctccct
     4741 aaatcctcag agagcactcc tttacctctg tgtccctccc aacctcagat ggactacaga
     4801 ggactgcaac cttgcctgcg gaccatgccc ctgtctgtgt gtccacccat ggctgagacg
     4861 gggtcctgct gcaccacaca cattgccaac cactcctacc taccactaag ctattggcag
     4921 cagccctgaa ggcagtcccc atgctactgc agacctatac attcctacac actaccttat
     4981 ccatcctcaa caccatccat tctgttgcca ccccactccc cctctggctt tataacactg
     5041 atcactccag atggctgctc acaaatccag agctctgtct ctgcagctgc ttggctcctc
     5101 tggctgttta tttttatttt tttaattttt tagacactgt ttattatata actctgatga
     5161 cctataactt gctatgtaga aacggctggc ctcaaactca cagaaatctg cctgctgtga
     5221 ctaaagttgc catgccctgt agctccaaac ccttgggggt ggggaaagtc tctgtttccc
     5281 tatctcctac aggctttaaa gagttcttca gtgagtcccc tagcttgctg ttaagggaac
     5341 tgtgcaagcc ttctaattat gagacccagt ggggtcttgc tgtcatctat ttatttgatc
     5401 tctcttggca tcctcctcca ccctgggtgc cacagtatca ccttccttcc tggttctgtt
     5461 cctgcctctc tgagcatact agcagcaaaa gagccaactc tggagctaga accaggttct
     5521 ggtcctgtcg ttcacacctg ctagctgtgc aaacttaggc aaatagcctt tgagcctttt
     5581 ccctctcatc tacgaagtgt ggataataac agaaccccat agagcagtgt gaacaattga
     5641 gaacacactg gctgggacag tcaatgctta aaatgcccca attaatgtcc taagtatttc
     5701 ttctcttcct attctggcct ttgttacatg atgaccatca gaaagttctt atctccagcc
     5761 cagacttcta tagatagatc ccacagtgtt ttctgtggat tttaacattt gtttcttgct
     5821 cctggatagt gagttgaaag gcaagggtta ggatctgtcc ttacccccca cccccacccc
     5881 agcatcttca atactatttc acgtatgata ggtggcagac ctccaataaa tatcgtctga
     5941 atg
//
