LOCUS       sequence-6859-ex        5321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  AUTHORS   Shoichet SA, Malik TH, Rothman JH, Shivdasani RA
  TITLE     Action of the Caenorhabditis elegans GATA factor END-1 in Xenopus 
            suggests that similar mechanisms initiate endoderm development in 
            ecdysozoa and vertebrates.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 Apr 11. 97(8):4076-81.
  PUBMED    10760276
REFERENCE   2  (bases 1 to 5321)
  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: "Proc Natl 
            Acad Sci U S A. 2000 Apr 11. 97(8):4076-81."
COMMENT     Sequence Label: p029 End-1::EnR
FEATURES             Location/Qualifiers
     source          1..5321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1368..1502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1392..1411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1446..1465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1619..1639)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1619..1637)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1658..1674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1658..1674)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1671..1693)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1682..1698
                     /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(1706..1712,1713..1730,1731..1736))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1751..1772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1889..1906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2060..2648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2140..2159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2819..3610,3611..3679))
                     /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     3442..3461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3680..3784)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3810..4265)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3947..3968)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4159..4178
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4337..5321
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccatcatg gactatcaac aacaggaaat cgaaaatatt
      121 ccaccagtct ctacaaatcg caaaattgtg aacaaaaagc cctcaacgtt ccacacgaac
      181 tctgtctgct ccaatccaaa ttgtagaact cgtgagacaa ctctatggag aagaactgat
      241 tcaggagcta ttgagtgcaa cggttgctcc ctgtacttcc gaaaaaatgg gattcaaaga
      301 ccagccgagt tgtgccgtaa aacaataatg aaaagaaacc gacgcccacg tgccgaagta
      361 caatctccaa caccagaaga ctcgaaacta tgtcataata ctactccact acagaacatt
      421 ccatcacaac atttctctct cgaggccctg gaggatcgct gcagcccaca gtcagcgccc
      481 agccccatta ccctacaaat gcagcatctt caccaccagc aacagcagca gcagcaacag
      541 cagcagcaaa tgcagcacct ccaccagctg cagcaactgc agcagttgca ccaacagcaa
      601 ctggccgccg gtgtcttcca ccatccggca atggccttcg atgccgctgc agccgccgct
      661 gctgcagctg ctgctgcggc cgcccacgct catgctgctg cactgcagca gcgcctcagt
      721 ggcagtggat cgcccgcatc ctgctccacg cccgcctcgt ccacgccgct gaccatcaag
      781 gaggaggaaa gcgactccgt gatcggtgac atgagtttcc acaatcagac gcacaccacc
      841 aacgaggagg aggaggcgga ggaggatgac gacattgatg tggatgtgga tgatacgtcg
      901 gcgggcggac gcctgccacc acccgcccac cagcagcagt cgacggccaa gccctcgctg
      961 gccttttcca tctccaacat cctgagcgat cgtttcggag atgtccagaa gcctggcaag
     1021 tcgattgaga accaggccag catattccgc cccttcgagg cgaatcgctc ccagactgcc
     1081 acgccctccg cctttacaag agtggatctg ctggagttta gccggcaaca gcaagctgcc
     1141 gccgcagccg ctactgcggc catgatgctg gaacgggcca acttccttaa ctgcttcaat
     1201 ccggctgcct atcccaggat acacgaggaa atcgtgcaga gtcgcctgcg caggagtgca
     1261 gccaatgccg tcatcccgcc gcccatgagc tccaagatga gcgatgccaa tccagagaaa
     1321 tctgctctgg gatcttaatc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1381 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1501 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1561 tttaattcgc ggcgcgccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc
     1621 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1681 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1741 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1801 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1861 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1921 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1981 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2461 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2581 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2641 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2701 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2761 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2821 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2881 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2941 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3001 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3061 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3121 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3181 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3241 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3301 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3361 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3421 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3481 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3541 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3601 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3661 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3721 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3781 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     3841 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3901 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3961 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4021 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4081 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4141 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4201 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4261 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4321 ttcgctatta cgccaggacc atagccaatt caatatggcg tatatggact catgccaatt
     4381 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
     4441 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
     4501 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
     4561 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
     4621 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
     4681 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
     4741 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
     4801 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
     4861 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
     4921 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
     4981 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
     5041 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
     5101 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
     5161 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     5221 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     5281 cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
