LOCUS       sequence-6860-ex        4923 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 4923)
  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 4923)
  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: p031 pCS2 End-1 ORF
FEATURES             Location/Qualifiers
     source          1..4923
                     /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    970..1104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(994..1013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1048..1067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1221..1241)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1221..1239)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1260..1276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1260..1276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1273..1295)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1284..1300
                     /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(1308..1314,1315..1332,1333..1338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1353..1374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1491..1508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1662..2250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1742..1761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2421..3212,3213..3281))
                     /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     3044..3063
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3282..3386)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3412..3867)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3549..3570)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3761..3780
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3939..4923
                     /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 atccaaccat gtatcaatcg tcttcttcat ctccatctcc
      121 ccatcttcca tatacctatg gagtcattga caaggatgga aatgttcatt cccatgaaat
      181 gcattttccg gataatcatg gaaatccttc gccactcgat gctacacaaa tgtaccaaca
      241 ggctccagtt caatatggtg gtggagatca atattttgat gattttgggc aatactttgt
      301 tcaatcgtac gatccagcac aacaatcgac accagtgcat ccaatgtttg aaagtctcga
      361 tatgatgaat tgctactcac accagtatcc ccaaattgga caagactatc ancaacagga
      421 aatcgaaaat attccaccag tctctacaaa tcgcaaaatt gtgaacaaaa agccctcaac
      481 gttccacacg aactctgtct gctccaatcc aaattgtaga actcgtgaga caactctatg
      541 gagaagaact gattcaggag ctattgagtg caacggttgc tccctgtact tccgaaaaaa
      601 tgggattcaa agaccagccg agttgtgccg taaaacaata atgaaaagaa accgacgccc
      661 acgtgccgaa gtacaatctc caacaccaga agactcgaaa ctatgtcata atactactcc
      721 actacagaac attccatcac aacatttctc ttaaatttat cttatctttt tatgtgtatg
      781 ctcctttgtt ccttttgatg atctctaaat gaatgtgaca gttctactga taagctcttt
      841 ttgtgtgcta ccaatcaggg gaactctaac taaagttttc tgtctctggg gatttctcta
      901 gtataaaaac attattaaag aattcaaggc ctctcgagcc tctagaacta tagtgagtcg
      961 tattacgtag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1021 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1081 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1141 cagggggagg tgtgggaggt tttttaattc gcggcgcgcc gcggcgccaa tgcattgggc
     1201 ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     1261 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1321 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1381 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1441 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1501 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1561 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1621 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1681 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1741 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1801 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1861 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1921 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1981 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2041 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2101 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2161 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2221 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2281 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2341 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2401 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2461 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2521 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2581 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2641 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2701 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2761 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2821 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2881 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2941 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3001 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3061 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3121 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3181 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3241 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3301 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3361 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     3421 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3481 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3541 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3601 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3661 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3721 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3781 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3841 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     3901 aagggcgatc ggtgcgggcc tcttcgctat tacgccagga ccatagccaa ttcaatatgg
     3961 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
     4021 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
     4081 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
     4141 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
     4201 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
     4261 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
     4321 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
     4381 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
     4441 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
     4501 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
     4561 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
     4621 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
     4681 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
     4741 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     4801 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     4861 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     4921 aac
//
