LOCUS       sequence-9965-ex        7276 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 7276)
  AUTHORS   Friedrich G, Soriano P
  TITLE     Promoter traps in embryonic stem cells: a genetic screen to identify
            and mutate developmental genes in mice.
  JOURNAL   Genes Dev. 1991 Sep . 5(9):1513-23.
  PUBMED    1653172
REFERENCE   2  (bases 1 to 7276)
  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: "Genes Dev. 
            1991 Sep . 5(9):1513-23."
COMMENT     Sequence Label: SA-betageo
FEATURES             Location/Qualifiers
     source          1..7276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(724..789,790..827)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     CDS             878..3919
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQ"
     CDS             3944..4747
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4007..4026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4617..4636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4779..4796)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4785..5009
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(5091..5109)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5130..5146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5130..5146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5143..5165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5154..5170
                     /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(5178..5184,5185..5202,5203..5208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5223..5244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5361..5378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5532..6120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5612..5631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6291..7082,7083..7151))
                     /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     6914..6933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7152..7256)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gcctcgaggc tagaactagt ggatcccccg ggctgcagat
      721 ctgtagggcg cagtagtcca gggtttcctt gatgatgtca tacttatcct gtcccttttt
      781 tttccacagc tcgcggttga ggacaaactc ttcgcggtct ttccagtggg gatcgacggt
      841 atcgataagc ttgatgatct gtgacatggc ggatcccgtc gttttacaac gtcgtgactg
      901 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
      961 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1021 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     1081 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     1141 tgcgcccatc tacaccaacg taacctatcc cattacggtc aatccgccgt ttgttcccac
     1201 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     1261 aggccagacg cgaattattt ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg
     1321 gcgctgggtc ggttacggcc aggacagtcg tttgccgtct gaatttgacc tgagcgcatt
     1381 tttacgcgcc ggagaaaacc gcctcgcggt gatggtgctg cgttggagtg acggcagtta
     1441 tctggaagat caggatatgt ggcggatgag cggcattttc cgtgacgtct cgttgctgca
     1501 taaaccgact acacaaatca gcgatttcca tgttgccact cgctttaatg atgatttcag
     1561 ccgcgctgta ctggaggctg aagttcagat gtgcggcgag ttgcgtgact acctacgggt
     1621 aacagtttct ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg
     1681 tgaaattatc gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga
     1741 aaacccgaaa ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca
     1801 caccgccgac ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg
     1861 gattgaaaat ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg
     1921 tcacgagcat catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatat
     1981 cctgctgatg aagcagaaca actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc
     2041 gctgtggtac acgctgtgcg accgctacgg cctgtatgtg gtggatgaag ccaatattga
     2101 aacccacggc atggtgccaa tgaatcgtct gaccgatgat ccgcgctggc taccggcgat
     2161 gagcgaacgc gtaacgcgaa tggtgcagcg cgatcgtaat cacccgagtg tgatcatctg
     2221 gtcgctgggg aatgaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa
     2281 atctgtcgat ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac
     2341 cgatattatt tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc
     2401 gaaatggtcc atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgatcctttg
     2461 cgaatacgcc cacgcgatgg gtaacagtct tggcggtttc gctaaatact ggcaggcgtt
     2521 tcgtcagtat ccccgtttac agggcggctt cgtctgggac tgggtggatc agtcgctgat
     2581 taaatatgat gaaaacggca acccgtggtc ggcttacggc ggtgattttg gcgatacgcc
     2641 gaacgatcgc cagttctgta tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc
     2701 gctgacggaa gcaaaacacc agcagcagtt tttccagttc cgtttatccg ggcaaaccat
     2761 cgaagtgacc agcgaatacc tgttccgtca tagcgataac gagctcctgc actggatggt
     2821 ggcgctggat ggtaagccgc tggcaagcgg tgaagtgcct ctggatgtcg ctccacaagg
     2881 taaacagttg attgaactgc ctgaactacc gcagccggag agcgccgggc aactctggct
     2941 cacagtacgc gtagtgcaac cgaacgcgac cgcatggtca gaagccgggc acatcagcgc
     3001 ctggcagcag tggcgtctgg cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc
     3061 catcccgcat ctgaccacca gcgaaatgga tttttgcatc gagctgggta ataagcgttg
     3121 gcaatttaac cgccagtcag gctttctttc acagatgtgg attggcgata aaaaacaact
     3181 gctgacgccg ctgcgcgatc agttcacccg tgcaccgctg gataacgaca ttggcgtaag
     3241 tgaagcgacc cgcattgacc ctaacgcctg ggtcgaacgc tggaaggcgg cgggccatta
     3301 ccaggccgaa gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat
     3361 tacgaccgct cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta
     3421 ccggattgat ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac
     3481 accgcatccg gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa
     3541 ctggctcgga ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga
     3601 ccgctgggat ctgccattgt cagacatgta taccccgtac gtcttcccga gcgaaaacgg
     3661 tctgcgctgc gggacgcgcg aattgaatta tggcccacac cagtggcgcg gcgacttcca
     3721 gttcaacatc agccgctaca gtcaacagca actgatggaa accagccatc gccatctgct
     3781 gcacgcggaa gaaggcacat ggctgaatat cgacggtttc catatgggga ttggtggcga
     3841 cgactcctgg agcccgtcag tatcggcgga attccagctg agcgccggtc gctaccatta
     3901 ccagttggtc tggtgtcagg ggatcccccg ggctgcagcc aatatgggat cggccattga
     3961 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4021 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4081 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4141 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4201 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4261 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4321 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4381 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4441 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgatga
     4501 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4561 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4621 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4681 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4741 cttctgaggg gatcaattct ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     4801 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4861 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4921 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4981 gcatgctggg gatgcggtgg gctctatggc ttctgagacg gaaagaacca gctggggctc
     5041 gatcctctag agtcgacctc gagggggggc ccggtaccca gcttttgttc cctttagtga
     5101 gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5161 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5221 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5281 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5341 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5401 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5461 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5521 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5581 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5641 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5701 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5761 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5821 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5881 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5941 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6001 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6061 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6121 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6181 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6241 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6301 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6361 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6421 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6481 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6541 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6601 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6661 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6721 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6781 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6841 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6901 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6961 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7021 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7081 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7141 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7201 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7261 tttccccgaa aagtgc
//
