LOCUS       sequence-388273-        5213 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 5213)
  AUTHORS   Shimamura K, Hartigan DJ, Martinez S, Puelles L, Rubenstein JL
  TITLE     Longitudinal organization of the anterior neural plate and neural 
            tube.
  JOURNAL   Development. 1995 Dec . 121(12):3923-33.
  PUBMED    8575293
REFERENCE   2  (bases 1 to 5213)
  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: 
            "Development. 1995 Dec . 121(12):3923-33."
COMMENT     Sequence Label: Nkx2.1
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2402..2420)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2441..2457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2441..2457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2454..2476)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2465..2481
                     /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(2489..2495,2496..2513,2514..2519))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2534..2555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2672..2689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2843..3431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2923..2942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3602..4393,4394..4462))
                     /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     4225..4244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4463..4567)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4594..5049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4681..4700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4891..4912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5176..5198
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5190..5207
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5191..5207
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggtc gacggtatcg ataagcttga tctttttttt
       61 tttttttttt tcctcctctt ccttcctcct ccagccgacg ccgaatcatg tcgatgagtc
      121 caaagcacac gactccgttc tcagtgtctg acatcttgag tcccctggag gaaagctaca
      181 agaaagtggg catggagggc ggcggcctcg gggctccgct cgcagcgtac agacagggcc
      241 aggcggcccc accggccgcg gccatgcagc agcacgccgt ggggcaccac ggcgccgtca
      301 ccgccgccta ccacatgacg gcggcggggg tgccccagct ctcgcactcc gccgtggggg
      361 gctactgcaa cggcaacctg ggcaacatga gcgagctgcc gccttaccag gacaccatgc
      421 ggaacagcgc ttcgggcccc ggatggtacg gcgccaaccc agacccgcgc ttccccgcca
      481 tctcccgctt catgggcccg gcgagcggca tgaatatgag cggcatgggc ggcctgggct
      541 cgctggggga cgtgagcaag aacatggccc cgctgcccag tgcgccccgc cggaagcgcc
      601 gggtgctctt ctcccaggcg caggtgtacg agctcgagcg acgtttcaag caacagaagt
      661 acctgtcggc gccggagcgc gagcatctgg ccagcatgat tcacctgaca cccacgcagg
      721 tcaagatctg gttccagaac caccgctaca agatgaagcg ccaggctaag gacaaggcgg
      781 cgcagcaaca actgcagcag gacagcggcg gcggcggagg cggcggtggc ggtgcgggat
      841 gcccgcagca gcagcaagct cagcagcagt cgccgcgccg ggtggccgtg ccggtcctag
      901 tcaaagacgg caaaccctgc caggcgggcg cccctgcccc gggagccgca agcctgcaaa
      961 gccacgcgca gcaacaagct cagcagcagg cgcaggcggc gcaagcggct gccgcggcca
     1021 tctcagtggg cagcggtggc gcgggtctag gagcacaccc aggccaccag ccgggcagcg
     1081 cagggcagtc cccggacctg gcgcaccacg cagccagccc cgcggggctg cagggccagg
     1141 tctccagcct atcccatctg aactcctcgg gctcggacta tggcgccatg tcttgttcta
     1201 ccttgcttta tggtcggacc tggtgagacc tgagatgcgc ttgagccccg cgcgacctca
     1261 acgcttcccc tctgccttcc gcaaagacca ccattcgccc gctgctccac gcgcttctac
     1321 cttttttaag aatctgttta tgtttagacc aaggaaaagt acacaaagac caaactgctg
     1381 gacgacttct tcttcttctt cttcttcttc ttcttcttct tcttcttctt cttcttcttc
     1441 ttcttcttct tcttcttctt cttcttcttc ttcttcttct tcttcttctt ctcctcctcc
     1501 tcctcctcct cctcctcttc ctccttcttg tccccgctcg ttcttttctt tctccccctc
     1561 ctcttctgtt tccttcttcc ttcatctttc cccccttcct ttctctttac tatctaaaac
     1621 ttgcagactt tttgtttttt aacataaaaa gaaaatagaa acagccaagc aaattcaacc
     1681 ctttacgaat tctttaaaca gagaaggaca gagaacaaat ttggggtgtc tttctggtag
     1741 ttcaaatggg ttcccaagct taggcatggc acagttttgg agcctgttct atgcttccat
     1801 ggccctgaac tctaaagacg gaaaactttt ctgtggatgc accctgccag caaagtgagc
     1861 ttgcttgtaa ataccaggat ttttcgtttg tttgtatgtt tcagaaggga ggacagacgc
     1921 tggagatagg aaagtcttca gcataaccca tttgtacctg acacaaagga agtgtcccct
     1981 cccaggcgcc ctctggccct acaggttcag tccaggctgg cctttcagaa aattgtttta
     2041 ggtttgatgt gaacttgtag ctgtaaaatg ctgttaaaag ttggactaaa tgcctagttt
     2101 ttagtaacct gtacattatg ttgtaaaaag aaccccagtc ccagtcccta gtccctcact
     2161 ttttcaaggg gcattgacaa acctgtgtat attatttggc agtttggtat ttgcagcacc
     2221 aatccttttt tttttttttt ctgttgtaac ttatgtagat atttggctta aatatagttc
     2281 ctaagaagct tctaataaat tatacgaatt aaaaagatgg tttttttcct gattaaaaaa
     2341 aaaaaaaaaa aaaaagcggc cgcggaggcc tctagacctc gaggagctcc agcttttgtt
     2401 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2461 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2521 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2581 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2641 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2701 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2761 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3241 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3361 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3421 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3481 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3541 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3601 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3661 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3721 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3781 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3841 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3901 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3961 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4021 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4081 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4141 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4201 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4261 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4321 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4381 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4441 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4501 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4561 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     4621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4801 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4861 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4921 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4981 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5041 ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     5101 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     5161 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttg
//
